For the submission: A tool for multiple targeted genome deletions that is precise, scar-free, and suitable for automation, Aubrey et al.
Figure S1. This gel demonstrates PCR1 products for the initial 11 ORFs in lanes 1-11 when a 10:1 ratio of primers (forward:reverse) was employed. Other lanes on this gel show PCR2 results, and are discussed later. Although amplification in PCR1 was good when equimolar amounts of the forward and reverse primers were used, the amplification product was sometimes weak or not visible when a 10:1 ratio of primers was employed. Alternative primer ratios might have proven effective, but this was not explored further. For example, when a 10:1 ratio of primers (forward:reverse) was employed in PCR1, only eight of the initial 11 ORFS amplified well. This can be seen in the lanes numbered 1-11. Lanes 2, 3 and 9 show poor amplification.
Figure S2. This gel shows the contrasting situation where equimolar amounts of primers are used for PCR1. The expected product was still obtained in PCR3 but sometimes with further amplification of the PCR1 and PCR2 products. Occasionally, even when PCR1 was successful at a 10:1 primer ratio, the PCR3 amplification produced further amplification of PCR1 and PCR2 products, presumably caused by failure of the SOEing reaction during the first 10 cycles of PCR3.
Figure S3. When testing the primers selected on the basis of these initial criteria, PCR2 failed for the ORF YGR125W and electrophoretic analysis indicated primer dimer formation that had not been predicted. This gel shows the problem for YGR125W and demonstrates that the reverse primer is at fault. The reverse primer is used in lanes 26, 27, 29 and all show far more dimer than the neighbouring lanes that did not have the reverse primer.
Figure S4. Replacement PCR2 primers were designed for YGR125W and YGL202W using the new improved method, and they gave the expected PCR2 product without apparent dimer production.
Figures S5a and S5b. In addition to products of the expected size (approx 1.25 kb), PCR2 gave an additional product approximately double the expected size (approx 2.5 kb). This additional product was seen on several gels, including the two given here.
Figure S6. Tests on PCR2 for YMR272C to vary the extension time showed that the larger product was still evident despite reduced extension time.
Figure S7. Tests on PCR2 for YMR272C excluding the primers (including just the forward primer (F), including just the reverse primer (R), including both primers (F+R), including no primers (NP) showed that both primers were needed to produce any product.
ORF ID | PCR1 product size | PCR2 product size | PCR3 product size |
YMR272C | 774 | 1266 | 1890 |
YDR093W | 680 | 1281 | 1796 |
YGL202W | 736 | 1457 | 1852 |
YHR163W | 592 | 1266 | 1708 |
YPR073C | 667 | 1266 | 1783 |
YGR125W | 595 | 1281 | 1711 |
YNL169C | 976 | 1266 | 2092 |
YJL134W | 567 | 1281 | 1683 |
YPL262W | 855 | 1281 | 1970 |
YDR074W | 981 | 1266 | 2097 |
YMR205C | 707 | 1266 | 1823 |
Table S1. Table of expected sizes of the products produced for the 11 ORFS.
ORF ID | PCR1 forward (primer 1) | PCR1 reverse (primer 2) | PCR2 forward (primer 3) | PCR2 reverse (primer 4) |
YMR272C | GCTTACATTTGCAGACGTCAAAAAACCTGACAAACC | CTTCGTACTAGTTTTAGCTTACCACCGCTTTTAGTG | GTGGTAAGCTAAAACTAGTACGAAGTTATTGGTCAGCCTCGTCCAAAATTGTCAACCTAGGAAAACACACATTACTTGCCTC | TTATTCATATTTCATTTTGGACAATGGGGCATCGGGGCCTAAGTAGATCCCAATACAACAGAT |
YDR093W | GTGATCATTTCTGTATAAGTTTTCTCAACTGAATATATAAATATATATCATTACTATGCT | ATATGTGTTGAGTTTATGCTATGATTCTTTTTAAAGAATTCTATTATTAAAAGGATTAGG | AATTCTTTAAAAAGAATCATAGCATAAACTCAACACATATTGATTACAATTTTTTTCTTCCTTTGTACTATGTATACCAACACACATTACTTGCCTC | TTAACGGTCCCTGCTGCGCTGGCTGAGCAGCGTCTCTGCATGATTGATCCCAATACAACAGAT |
YGL202W | AAGACATTAACATCATTTTTCGTGCTGTTAGGGTGGTG | GATAGTAACGATCGGTTGTCCTATTCAATGTCTGTATC | AATAGGACAACCGATCGTTACTATCAGAGGTAAATACGTTGGAAGAATATATATAAGGTGAAAAACACACATTACTTGCCTC | CTATTTGGAAATACCAAATTCTTCGTATAAAGTATCACCTAATCTGATCCCAATACAACAGAT |
YHR163W | CAAGACGGTGCATTTCTGGGCTCCTACTTTGAAATGGGGTCTGG | TTGTCGCGGACCAGGGGTAAAGCAATTGATGGTGCATTGCCTTC | TTGCTTTACCCCTGGTCCGCGACAAAAAAGACACACATGCGAGCTTTCGAACCTCAGATGCTAATCACACATTACTTGCCTC | CTAAAAAGTTTTCGTTTGAACTTTTCCAAAAGCTTCCTCATTAACGATCCCAATACAACAGAT |
YPR073C | TTGTTTCTCTACCAGATTTCTTCACTTTTGACAAACAACCCCTAGAATACAGACTATC | TTTCGTGTTAACTTTCCCTTCTCAGTTTTCTATCGCTTATCAAAAATCACAGGGTTTC | CTGAGAAGGGAAAGTTAACACGAAAGCTAGCAAGTAATGATGGTGCGGTCTACATCGAGTAAAAACACACATTACTTGCCTC | TTATAACTCTTTTTTTAAAAACTGTTTGGAGAAATAGGTGATTTGGATCCCAATACAACAGAT |
YGR125W | AATCTATATAGATATATGTATATAATAAAAGTTTTCAATCTGCTGACC | ATTTCTAAATTAAATTTTTGAGATTTTAAACTGTCTTGCTTATAGACC | AGCAAGACAGTTTAAAATCTCAAAAATTTAATTTAGAAATGAGCGTACACGTATATTTATTTAATTTCTTTTATTGCCGGCACACATTACTTGCCTC | TCATGCGCTAACAAGTGTGTAACCTAGTAATTCTTTGAATCTGGTGATCCCAATACAACAGAT |
YNL169C | ACCACTAGACCTGCAAACTCGACTTTCAATGGTTTAAATGAAG | GCTGGCTTTGCTTTTCCTTCTTCTTCAAAAAATAACGACCAAG | AAGAAGAAGGAAAAGCAAAGCCAGCAAGCAATCATATGTAAAGTTAGCATTTATTTTGCTGTATACACACATTACTTGCCTC | TCATTTTAAATCATTCTTTCCAATTATGCCTAATTTCTGTCCCATGATCCCAATACAACAGAT |
YJL134W | CACATTGTCGTCAAAAGGTAATCTCCCCAGAAGTTTTCACTC | TTTTTATCTATACAAATGGCTTGTGTCCGCTGGTCTGGTAGG | TACCAGACCAGCGGACACAAGCCATTTGTATAGATAAAAAAACAAAAGGAACAAAAAAAAAAAAATAGACAAAATGTAGTCACACATTACTTGCCTC | TTATGCTATATTTAAGAGGGAAAATAGGACGGGGCTGCACATTACGATCCCAATACAACAGAT |
YPL262W | TTATCCTTCTTATTCTTCTTTGCCATTAATCACTTCTAC | GATTGTATCTCTTATCCGTTTAATAGTTAGACTTTATGG | TCCATAAAGTCTAACTATTAAACGGATAAGAGATACAATCACGAGCTAAATACCTAATAATATACAAGTTTTTTATGTCTCACACATTACTTGCCTC | TTATTTAGGACCTAGCATGTGTTCAGGAACAACCCATTCATCAAAGATCCCAATACAACAGAT |
YDR074W | CTATCTTCTCTTGGCAAGAGTGTTCGAGATATGGTAG | TTCGGCACAGAAATAGTGACAGGCAGTGTTATTTTGG | TGCCTGTCACTATTTCTGTGCCGAAAACACCCTTTTTAACGAAATGGTTATGACTAGACAGACATCACACATTACTTGCCTC | TCAAACCTTTGCGCCGGTGTAAGAAGCCGATCTTTTCATAACATAGATCCCAATACAACAGAT |
YMR205C | CCATCTCCACGCTAAAAGGAAAACGAAGATTAAGATAAAGTTGGG | TGCGTATGGTTAGTTCTTGGCCAATGCTAAACTAGTCTCTAAATC | ATTGGCCAAGAACTAACCATACGCAAAGAAAATGACCTTTTATTACACTTTCTATTATTAATGTCCACACATTACTTGCCTC | TTAATCAACTCTCTTTCTTCCAACCAAATGGTCAGCAATGAGTCTGATCCCAATACAACAGAT |
Table S2. Table of primer sequences for the 11 ORFs
ORF ID | PCR2 forward (primer 3) |
YGL202W | AATAGGACAACCGATCGTTACTATCAGAGGTAAATACGTTGGAAGAATATATATAAGGTGAAAAAACACTCCTCAGAAGCTC |
YGR125W | TCATGCGCTAACAAGTGTGTAACCTAGTAATTCTTTGAATCTGGTCTAGAGATCCCAATACAAC |
Table S3. Further forward primers were then chosen for PCR2 for YGL202W and YGR125W
ORF ID | CPS A to CPS D with gene | CPS A to CPS D gene deleted | CPS A to RinklURA3 | CPS D to FinklURA3 |
YMR272C | 1801 | 646 | 709 | 1198 |
YDR093W | 5460 | 621 | 659 | 1223 |
YGL202W | 2240 | 737 | 744 | 1254 |
YHR163W | 1410 | 660 | 769 | 1152 |
YPR073C | 1041 | 555 | 651 | 1165 |
YGR125W | 3725 | 614 | 651 | 1224 |
YNL169C | 2051 | 548 | 650 | 1159 |
YJL134W | 1785 | 555 | 660 | 1156 |
YPL262W | 2167 | 700 | 766 | 1195 |
YDR074W | 3393 | 702 | 667 | 1296 |
YMR205C | 3196 | 316 | 552 | 1025 |
Table S4. Expected product sizes using the confirmation primers. The four resultant strains were verified using confirmation primers from the EuroFAN project.
Figure S8. Confirmation gel for strains YHR163W, YPR073C, YGR125W, YJL134W after selection on FOA, with expected product sizes shown at bottom of lanes.
Figure S9. Confirmation gel for strains YGR125W and YNL169C after selection on FOA, with expected product sizes shown at top of lanes.
ORF | Raw sequencing data files |
YHR163W | 4F, 4R |
YPR073C | 5_1F, 5_1R, 5_2F, 5_2R |
YNL169C | 7_2F, 7_2R, 7_4F, 7_4R |
YJL134W | 8F, 8R |
Table S5. Sequence results for the four strains with confirmed deletions.
ORF | Alignment files |
YHR163W | YHR163Wdeletant.txt |
YPR073C | YPR073Cdeletant.txt |
YNL169C | YNL169Cdeletant.txt |
YJL134W | YJL134Wdeletant.txt |
Table S6. Alignments of sequencing results with expected sequence after the deletion.
Well | Strain | Reason chosen | PCR1 notes | PCR2 notes | PCR3 notes | Colonies on URA- |
A1 | YAL023C | 32 good | 273 | |||
A2 | YAL056W | 32 good | 168 | |||
A3 | YBL069W | 32 good | weak | 30 | ||
A4 | YBR052C | 32 good | 200 | |||
A5 | YCL063W | 32 good | weak | 9 | ||
A6 | YCR095C | 32 good | wrong size | weak | 84 | |
A7 | YDL174C | 32 good | 100+ | |||
A8 | YDL191W | 32 good | 22 | |||
A9 | YDR222W | 32 good | 100+ | |||
A10 | YER122C | 32 good | no visible product, some dimer | 12 | ||
A11 | YER158C | 32 good | wrong size | no visible product, lots of the PCR1 wrong size product | 0 | |
A12 | YFL001W | 32 good | 18 | |||
B1 | YGL254W | 32 good | weak | 20 | ||
B2 | YGR163W | 32 good | weak | 140 | ||
B3 | YGR241C | 32 good | 13 | |||
B4 | YIR013C | 32 good | 170 | |||
B5 | YJL065C | 32 good | 120 | |||
B6 | YJR153W | 32 good | 210 | |||
B7 | YKL132C | 32 good | 172 | |||
B8 | YKL191W | 32 good | 132 | |||
B9 | YLR065C | 32 good | weak? | 72 | ||
B10 | YLR079W | 32 good | no visible product, some PCR1 | 2 | ||
B11 | YLR091W | 32 good | no visible product | no visible product | no visible product, some PCR1 | 0 |
B12 | YML097C | 32 good | no visible product, some dimer | no visible product, some dimer | 24 | |
C1 | YMR035W | 32 good | 240 | |||
C2 | YMR103C | 32 good | 106 | |||
C3 | YNL129W | 32 good | 200+ | |||
C4 | YNR057C | 32 good | no visible product, made PCR1 | 4 | ||
C5 | YOR076C | 32 good | weak | 72 | ||
C6 | YOR106W | 32 good | 68 | |||
C7 | YPL202C | 32 good | 208 | |||
C8 | YPR054W | 32 good | no visible product, produced a small product | 13 | ||
C9 | YAL058W | GC content, 3prime 0,1, overall 37.29 | no visible product | 44 | ||
C10 | YAR028W | GC content, 3prime 2,3, overall 35.42 | no visible product, some small product | 12 | ||
C11 | YBL033C | GC content, 3prime 4,4, overall 40.00 | weak | no visible product, some PCR2 | 21 | |
C12 | YBL055C | GC content, 3prime 2,4, overall 43.14 | weak, dimer | no visible product, some PCR2 | 52 | |
D1 | YBL091C | GC content, 3prime 0,2, overall 35.71 | 432 | |||
D2 | YBR077C | GC content, 3prime 5,5, overall 38.24 | weak | no visible product, dimer | 12 | |
D3 | YCL050C | GC content, 3prime 3,4, overall 21.43 | no visible product | 500+ | ||
D4 | YDR071C | GC content, 3prime 3,5, overall 24.49 | no visible product | no visible product | 1 | |
D5 | YDR330W | GC content, 3prime 3,3, overall 36.73 | weak | 105 | ||
D6 | YER115C | GC content, 3prime 2,6, overall 33.33 | no visible product | no visible product | 0 | |
D7 | YHL044W | GC content, 3prime 2,2, overall 42.59 | 440 | |||
D8 | YHR207C | GC content, 3prime 7,5, overall 39.02 | no visible product | weak | 29 | |
D9 | YIL079C | GC content, 3prime 3,3, overall 44.00 | weak plus dimer | weak | weak?, produced PCR2 | 50 |
D10 | YIL111W | GC content, 3prime 4,4, overall 37.84 | no visible product, some PCR2, some small product | 112 | ||
D11 | YJL058C | GC content, 3prime 4,5, overall 37.25 | 154 | |||
D12 | YJL128C | GC content, 3prime 1,4, overall 30.00 | no visible product | weak product, some PCR2 | 32 | |
E1 | YKL023W | GC content, 3prime 2,5, overall 57.14 | 32 | |||
E2 | YKL166C | GC content, 3prime 2,2, overall 43.75 | weak | no visible product, made PCR1 | 9 | |
E3 | YKR039W | GC content, 3prime 4,5, overall 21.95 | weak | 114 | ||
E4 | YLR218C | GC content, 3prime 6,6, overall 48.39 | weak | 153 | ||
E5 | YLR255C | GC content, 3prime 2,4, overall 60.00 | weak | 81 | ||
E6 | YLR350W | GC content, 3prime 7,5, overall 33.33 | 239 | |||
E7 | YML048W | GC content, 3prime 4,6, overall 53.85 | 213 | |||
E8 | YML088W | GC content, 3prime 3,4, overall 45.10 | no visible product | no visible product, lots PCR2 | 249 | |
E9 | YMR018W | GC content, 3prime 5,5, overall 38.64 | no visible product, lots dimer | weak, lots PCR1 and some small product about 600 | 44 | |
E10 | YMR278W | GC content, 3prime 1,2, overall 39.22 | no visible product, dimer | 53 | ||
E11 | YNR058W | GC content, 3prime 6,6, overall 44.44 | 332 | |||
E12 | YOL158C | GC content, 3prime 4,5, overall 40.00 | no visible product | no visible product, some PCR2 | 12 | |
F1 | YOR008C | GC content, 3prime 2,5, overall 27.27 | 42 | |||
F2 | YPR037C | GC content, 3prime 3,4, overall 51.35 | weak | 201 | ||
F3 | YPR040W | GC content, 3prime 2,4, overall 47.73 | 152 | |||
F4 | YPR051W | GC content, 3prime 1,2, overall 45.24 | 36 | |||
F5 | YBR273C | self dimer tests | 492 | |||
F6 | YAL030W | self dimer tests | 736 | |||
F7 | YAL065C | self dimer tests | wrong size? | 512 | ||
F8 | YAR002W | self dimer tests | no visible product | no visible product, some PCR2 | 9 | |
F9 | YBL049W | self dimer tests | weak, some small product | 69 | ||
F10 | YBR144C | self dimer tests | two weak bands | no visible product, lots PCR2 | 320 | |
F11 | YBR185C | self dimer tests | weak | 135 | ||
F12 | YBR255W | self dimer tests | 113 | |||
G1 | YCL044C | self dimer tests | ok, but dimer present | no visible product, dimer | 29 | |
G2 | YCL047C | self dimer tests | 294 | |||
G3 | YBR268W | self dimer tests | 445 | |||
G4 | YAL002W | dimer tests | 77 | |||
G5 | YBL082C | dimer tests | no visible product | 160 | ||
G6 | YCR045C | dimer tests | 500+ | |||
G7 | YDL106C | dimer tests | 500+ | |||
G8 | YIL148W | dimer tests | no visible product | no visible product, lots PCR2 | 42 | |
G9 | YJR073C | dimer tests | no visible product, lots dimer | no visible product, some PCR2, some small product | 100 | |
G10 | YLR011W | dimer tests | weak | 14 | ||
G11 | YCL044C | dimer tests | weak | 101 | ||
G12 | YDL022W | dimer tests | weak | 4 | ||
H1 | YDL175C | dimer tests | no visible product | 121 | ||
H2 | YGL211W | dimer tests | weak | no visible product, made PCR1 | 72 | |
H3 | YAL009W | pcr2 dimer tests | 259 | |||
H4 | YBR185C | pcr2 dimer tests | 824 | |||
H5 | YBR270C | pcr2 dimer tests | weak | no visible product | 117 | |
H6 | YCR025C | pcr2 dimer tests | 1004 | |||
H7 | YDL012C | pcr2 dimer tests | weak | 72 | ||
H8 | YDR375C | pcr2 dimer tests | no visible product, also lots small product | 70 | ||
H9 | YLR035C | pcr2 dimer tests | weak? | 67 | ||
H10 | YLR412W | pcr2 dimer tests | multiple product | 500+ | ||
H11 | YML089C | pcr2 dimer tests | 140 | |||
H12 | YPL033C | pcr2 dimer tests | no visible product, dimer | weak | 18 |
Table S7. Table of properties of the PCR results for the 96 ORFs.
Choices of ORFs were determined by the following:
The 32 ORFS chosen to have good primers:
The 32 ORFS chosen to test dimerism scoring:
The 32 ORFS chosen to test effect of GC content:
|
Algorithm S1. Summary of reasons/methods for choosing the 96 ORFS.
ORF ID | Sequences of forward and reverse primers | Percent G+C | G+C at 3' end | Commenta |
YCL050C | CTTTTTTTTATTGTGTTGGTTTTTATATGTTTTGTTATGTATTGTTTATTTTCCCT TTTTATTTTAATTTTTACTTTTCTGTTTGTTCTAAAATCTATCTAAACTGGCTTTC | 21.4 (12/56) | 3/8 4/8 | Required annealing at 55 ° C |
YKR039W | TTATTTATTTTTTTTTATACCATTTCCTTTTGATAAGGGGT TTTTTATTTCTTTTTTTTGTTTCTTATAAATGTTGCTGTCC | 22 (9/41) | 4/8 5/8 | Weak product |
YDR071C | TTTGTTTTACAACCAAAAGTTTTAAAAAAGTGGATTCATATAAGGTTAG TTTAACTTTTTATTTAATCGTAATGTATTGGTGTTATTTTTTCTCCGAG | 24.5 (12/49) | 3/8 5/8 | Required annealing at 55 ° C |
YOR008C | AAATAATGTGGAGATCTTATCTTGTTTTTCTCTTCTTTATGACA TATTTAAATAGAATTTTTTATCCTGAGTACTAAATCAGCCAACG | 27.3 (12/44) | 2/8 5/8 | |
YJL128C | GTAGTATAGTATTGCAACAGGAAAAAAAAATCTTATTGCGTAGCACTCTGTATCTATTTT CTTTGAGAAATTAACGTCTCGATCTGCTTAATATAATAATGTATCTATAGAAATGTACGG | 28.8 (17/59) | 1/8 4/8 | Required annealing at 55 ° C |
YER115C | AATAGTAAGGAACAAACTGTTGATTTAACTGCAGATGCCATAGACTTA ATACAAGTTTGAACAAAATACAACTCAAAGTAAAATGGTAGCTGCACG | 33.3 (16/48) | 2/8 6/8 | |
YAR028W | GTTGTAAACCGCTAATGATTATGTTCCTAATACCTGTGTGTTTGGTTT GGTCAATTGATATCTTGCCTTAATATGGCACTTATAGATGTTATCCTG | 35.4 (17/48) | 2/8 3/8 | |
YJL058C | CGGAAGTGAATGAGAATACCAACAGTTTTACTGATGATTATATTGACGGAG AATGAAAGAACTTTATTTCTACAGAGGTCAATTGGCGTTTCTGTCACCTAG | 37.3 (19/51) | 5/8 4/8 | |
YOL158C | AGAATCATCTCTACTACACACTGCATTATGAACTGCAGCCACATTTATCTAAGGG TTTCAGAGAGACATTCACGAGCACTTTTTTGCTATATTAGAGTTCTGCTAGGACC | 40 (22/55) | 4/8 5/8 | Required annealing at 55 ° C |
YBL055C | GAGAAAGTGAGTGGGAATGGACAAGATTATTTGGGTGAGGGTGTTTGCTTA ATAACTATAACAGTACTGGCGCCTCAAAAAAATGCCTTGGCAGCGAAAGTG | 43.1 (22/51) | 2/8 4/8 | |
YML088W | CTCTGAAGAGTTTAGCGGGCTCTCAGACGGTTTTTTCGCTTTTGGCATTTT GCTGTCTTATTCCCTAGTGTCGGCCACCTGATAAAGGTTTTTAACTCGTAC | 45.1 (23/51) | 3/8 4/8 | Required annealing at 55 ° Cb |
YPR040W | GCCGTCCTGCTAGATGTCTGTACGGGATACTTCTCAATCAACTT TTTTTCTGGGGAGCTGTTGTGTCTAAAGCTGCCCTTAGGTCATG | 47.7 (21/44) | 2/8 4/8 | |
YPR037C | TGCCCTTATACCGCGTAGTGTAGGTCCAGTCGTACTT CCCTTCGTTCCTTGCACAATCTGTCCAGACTTTCCTC | 51.4 (19/37) | 3/8 4/8 | |
YML048W | GCTTTTGCCCTTGACCCTGAATGCGGCAGATCCAGCGACTTCATCGAGGTTA GCTTGCAATCGCCCTCTATCAAAACAAAGTCCACCGATCTCTGCTGCCCCTC | 53.8 (28/52) | 4/8 6/8 | |
YKL023W | TTCCCCATATTGGGAACCTGCTCTGCCCTGCTCAGCACGTAG GCTTGCACGTTATACCTGGCCGGGATTGGGGCGCTATTCTTG | 57.1 (24/42) | 5/8 2/8 | |
YLR255C | AGGGTGGAGAGTTAGAGCTGGTGTTGGCACCGGGCGTAGTTGCTG TCACCCGTCGGGCGAAATCCACCCTCGCGGCCTCGTTTAGAATAC | 60 (27/45) | 4/8 2/8 | |
Table S8. Results of investigation into overall GC content. a Where there is no comment, the expected amplicon was obtained using the standard annealing temperature of 58 ° C (See Methods). b Only 6 of the 21 bases at the 3' end of the forward primer are G or C. In general, reactions involving primers with an overall G+C content below 30% either gave a weak product or no visible product at all. However, this was not always the case. Furthermore, some primer pairs with higher G+C contents also failed to yield a visible product. In spite of the apparent failure of some of these reactions under standard conditions, all primer pairs chosen to test overall G+C content yielded the expected product when the reduced temperature touchdown PCR was employed (touchdown PCR with annealing from 60-55 ° C for 5 cycles and then 55 ° C for a further 25 cycles). We do not advocate the use of a lower annealing temperature for all ORFs since stringency is reduced. We also examined the efficacy of amplification in relation to the frequency of G+C residues at the 3' extremity of primers with an overall G+C content above the 30% threshold (Supplementary information Table S9). We expected reactions to fail if primers lacked a GC clamp and contained two or less G+C residues among the last 8 at the 3' end. In fact, some primers of this nature did generate amplicons under standard annealing conditions (58 ° C) whereas others did not.
ORF ID | Sequences of forward and reverse primers | Percent G+C | G+C at 3' end | Commenta |
YLR350W | ACGATTATGTATATCCATATATATATATACACTACAAGCCGACCG GATGAAACTGTTTCTTATGCGTTTTTATAATGGTATAGTCTTGCG | 33.3 (15/45) | 7/8
5/8 | |
YHR207C | AAGGATAATAACAGAATTTAATGGCTTGATATGGACGGGCG TTTTGTGCCACTTTTCGTTATTTTGTATCAGTGCACGCTTG | 39.0 (16/41) | 7/8
5/8 | Required annealing at 55 ° Cb |
YNR058W | TAGTACATTGTGCAACCTGTGCCATAATATTGACACCGAACTTCTTCAGGGCCT TAGTTGTTATCCTAGCAATAGAAAACGAATTGCGCTGTTGACATCCTCCCGACC | 44.4 (24/54) | 6/8
6/8 | |
YLR218C | GTAGTCTTTACGGTAATGGAGTTGGCCCTTG AAATGAAGCGAATGGATGAGAAGAGGGGAGG | 48.4 (15/31) | 6/8
6/8 | |
YBR077C | CGTAGGTGTTTGACATTGAATATTTATACCGGCT ATTGCTTCAATATTTTGGATACCGTTTGCTGCTG | 38.2 (13/34) | 5/8
5/8 | |
YMR018W | AATTATATTCTGCGCTTGCCTTTTATATTAAGCGTCACAACGCC CGTGCTTACAAAAGGTACCCTTTATTCTTAATGATTCTCCACTC | 38.6 (17/44) | 5/8
5/8 | Required annealing at 55 ° C |
YBL033C | GGTTCAAAAAGTGGAAGTAGTAGTGCTCTT CCTGTAGGTTTGTTGTAGTTTTATGTACGC | 35.4 (12/30) | 4/8
4/8 | |
YIL111W | ATGATATTGTTGAAGGATATCGACTTGCTCTCAAAGG GCTCTTTACAATGAATATGTGGCGTTATTCCATTGTG | 37.8 (14/37) | 4/8
4/8 | |
YDR330W | CACGATGTAACCCAGTTATAGAACCTATTCTGTAGTTTGTTGCAGTTTT ATAAAAGGTATGTTATTATTGCCACACGTCGTAGTCTATGCGTTTGTAG | 36.7 (18/49) | 3/8
3/8 | |
YIL079C | CTTTCCTCGATGAAAGTTGGTTCAGGATCCAAGTACAAGGACTGTTGCTT AATATCTTAGCTCCGTGGTCCGCTTCCTTTGTATTCCTCATGTGAGAATG | 44.0 (22/50) | 3/8
3/8 | |
YHL044W | GCAGTACTACAACTCTGTGCGGTGGGAAAAATTTATTCTTCGAGGCACAGTATT AGCCTTAGCGGGCCCTCACTTGCTTTTGAATTTGTCTTTATACCACAAGTTAAC | 42.6 (23/54) | 2/8
2/8 | |
YKL166C | CCAGGTACGAGTGATTTAGGAAGGTCTGAAGAAAAGCGACTATGCATT TTTGTGCAGGCTCGCTCTTTCCTTGTACAACCACCGATATACAATATG | 43.8 (21/48) | 2/8
2/8 | |
YPR051W | CAACCAAGGTTCGTCGTCAGTAAGCTAGTTACGGAATGGTAA GTCCTGACCCAACGCCTTTTCTGTAGTAGTGGCTTTTTTATC | 45.2 (19/42) | 1/8
2/8 | |
YMR278W | TTTAATCACCAAGAACCCTAGCACTTATAAAGCGCGTCTAGCGAAAAAGTT TTTTTGAAATTGCAGCAGCTAATGGCTCGAGAGTACTGTTGAGTATTTAGG | 39.2 (20/51) | 1/8
2/8 | Required annealing at 55 ° C |
YBL091C | TGCTTACTTTTCGTTTCCACTTTATCTTGCTGTTCGTTCGAGCAACAGTATTTATT TTTTCAATACGGTAGAGCTTCTACAGTACTTGTTGATGTAAAAGCCCACTTATTAG | 35.7 (20/56) | 0/8
2/8 | |
YAL058W | CAAGTACATTCAAAATACAAGAAAGGCTGGCTGGGATAGCACCAATGAGACTATAAAAA AGTACTACTTTGTGTGGCCGTTATCTATATTAGAAATGCGTTGCCGTTCACTTTTTTAC | 37.3 (22/59) | 0/8
1/8 | Required annealing at 55 ° C |
Table S9. Results of investigation into G+C content of the 8 bases at the 3' end. a Where there is no comment, the expected amplicon was obtained using the standard annealing temperature of 58 ° C (See Methods). b This reaction gave a product of unexpected size. See also Table S8.
ORF ID | PCR1 forward (primer 1) | PCR1 reverse (primer 2) | PCR2 forward (primer 3) | PCR2 reverse (primer 4) |
YAL023C | TTCCTGTATTTGCGTCTGCCACCTACGTGA | GATTGCTGGACCACGGTTCGAAACAGAATG | TGTTTCGAACCGTGGTCCAGCAATCATTAAGATCATCGTAGTAAGTACATAAGTAAAAAACAAGACACACATTACTTGCCTC | TCATGCTTCTTGCTTGTCGGCAATGTCCCAAGTGGAAAACCAGTTGATCCCAATACAACAGAT |
YAL056W | AGGCGCAAATTATTGGGAATAAGACTTGGCTG | GCCTATGATTTAGTAATGCGACAATGGACCTG | TTCATTGGCAGGTCCATTGTCGCATTACTAAATCATAGGCAAAGGCAGTTTTCAATGCTTTCACTTTGTAAACTTTGTTTCACACATTACTTGCCTC | TTATGCACTAGGATTTACGCTAGGAAAAATACTTTTCGTTATGGTGATCCCAATACAACAGAT |
YBL069W | CCTTGTCCCTGTGGAGATAGGTTTCAAATATATCTGGATGACATGTTTGAGGGCGA | TGTTCAACGTTTGCCTCTTGGTATGCAATGTCGTATTTGTAGCTTCCGGAATGCTC | CATACCAAGAGGCAAACGTTGAACAATACCCACTTACATTTACAATATGTTTTATTTTTAAACTTAGCGACAAGAAGAGATAG | CTAGAATTTTTCGACGTTCATGATTAACTTTCCTTGGGCACGTTGGATCCCAATACAACAGAT |
YBR052C | TGTTTGCTTCGTGTTGACCTATTAGCTCACAAGATAGAGCTTGTTTGGAA | TATCTATGTGGCGTAGTATGTGCTTATAATAGTGAGTAGTATGCTGGAGG | AAGCACATACTACGCCACATAGATAAACCGTATATATCTTTACATAAGTACGATATGGTATAACTCACACATTACTTGCCTC | TTACAAATCTTTGATACGGTCGTAAAAGGCCTTCCCTTGAAGTTGGATCCCAATACAACAGAT |
YCL063W | TAGCGAAAACCTGAGGAGATCTTTTACCGTCCTT | TAATCGATGTGTCCTTGTTTCCTTATGCGAGCTG | AAGAAACAGCTCGCATAAGGAAACAAGGACACATCGATTATTAATGCCTCCTTTACCTAACCTACTCTTCTTTTGCTCCAAGCGACAAGAAGAGATAG | TTAAAACAGCAGTTCTGTATTCAAAGCATCTTGCAATTCTTCTATGATCCCAATACAACAGAT |
YCR095C | ATCCTTTGTTCACTTTCTCTTTCTGCACCTACATCACCGTC | TATGAATTCAGGTTATGGATCGCACCAGTGCTGTAAGTGTC | GTGCGATCCATAACCTGAATTCATATTTACATTTTTACGTATATGTATAGATGATTTCATGATAACACACATTACTTGCCTC | CTATGTAAAAATATGTTCCCTATAAAAGTGATGCTCTTCCGGAACGATCCCAATACAACAGAT |
YDL174C | ATAGTTGATATTGTCGTTAGAGTCCAGTTGTTGGGCG | TGTTCTTGCTTTTGTTTCCAGCTGGATCGGAATGATG | CCAGCTGGAAACAAAAGCAAGAACAGTATCTGATTTTCCTTTTTCACCCTTCACGTAAACCTGAACACACATTACTTGCCTC | TCACCTGTAATCATTAGCGGGCTCGTTTGGATCAGTTTTAAAGATGATCCCAATACAACAGAT |
YDL191W | TACAACGCATTTTCATTTGTTTTACAGCACCCCTG | TTCGAATATACTGTCTCACTGTACCTATTCTACGG | GGTACAGTGAGACAGTATATTCGAAACAAAATAATAAAGACTGACAACTGCAGAACAGAGAGGATCACACATTACTTGCCTC | TCATCTCAAACATACCATTTCGAATATACTGTCTCACTGTACCTAGATCCCAATACAACAGAT |
YDR222W | CAAGACTTTCGAATACGAGCCCCCTAAGTTCAATTTAAGTGAATGACTGGAA | CACTAAATGAGTCTCTATGTATGTACGATGTATCTTATACGCCCTCACTGTC | GTATAAGATACATCGTACATACATAGAGACTCATTTAGTGACAAAAGCGTGAACACTTCCACATGAAATTCGTTTTTTGTCACACATTACTTGCCTC | TCAACTAATGAAGGTGCTCTCAACGATAGAAATACCTTTCTCGTGGATCCCAATACAACAGAT |
YER122C | TGCGCTTGACAAGAATGAAGAATGTTCTCTCCCTCAAA | TGTCGCTATGTGTATACTAAATTGCTCAAGAAGACCGG | ATCCGGTCTTCTTGAGCAATTTAGTATACACATAGCGACAGTTTAATTTTCTCTTTAATTGAATTTTTTTTTTTTCATTGCACACATTACTTGCCTC | TTATTTTCTTAAGTAGTCCCTTAAATAACTACCGAGTTTTTCAGCGATCCCAATACAACAGAT |
YER158C | ATGCGTAGAAGAAGGCCAAACACAACCGGAGGAA | GCTGATTAAGAGTGAGTGGAAGTAGTAGCGAGCG | CTACTTCCACTCACTCTTAATCAGCCGGGTTCATCAAGCTTTTTTCCTTCTTGAGACTCTGTTGTCACACATTACTTGCCTC | TTATAAGTAGTAATGTGTATACTTAGCGCTATCCTGGTGAACGCGGATCCCAATACAACAGAT |
YFL001W | TACGACACAGTAATTTAATTTCAAAGAGGAGGGCGCCTGG | TCAAGGGAGGTTATAGTAGGGCAGTAAAGATTGCATGTGG | ACTGCCCTACTATAACCTCCCTTGAAATAATATATAAACCTGTATAATATAACCTTGAAGACTATCACACATTACTTGCCTC | TTACTTATTTTTGTTGTTCTTTTTCTTGGAGTATTTAGCATTAACGATCCCAATACAACAGAT |
YGL254W | AGTGTTGGTGATGAGTTCTCCTCCCTATACCCT | TGTTTTGGTGGCCACGTATTCTGGTACCACTTG | ACCAGAATACGTGGCCACCAAAACAACGCCATAGAAGAGCAATTTCCGTCCTCTACTCATGTGATAGCGACAAGAAGAGATAG | TCAGTATTCGAATAAATCCCAGACGCTAGGCTTATATAAATGAATGATCCCAATACAACAGAT |
YGR163W | AACAGATTCATGAGGAAGAGGAGAGACAACGTCAATT | GTTGTATGTGTATTAGTACCGTTGTCCTGGAGTTTTG | TAACAAAACTCCAGGACAACGGTACTAATACACATACAACAAGACGTAAGGCATGAAAATATTAGGGTATATAGATACATACAATCATATGGGAGAAG | TCACACATCGTCCTCAATACTATTTTTAGCTTGACTGGCTCTTGCGATCCCAATACAACAGAT |
YGR241C | TTCTCCAATACTCCAATTGCAACGCACCATATTATCCGGGAGAATACACG | TGTTTCACAAACACGCCTCTATATCTATTATAGGGGTAGCCGAGACAGTC | GATATAGAGGCGTGTTTGTGAAACATGAGCGATAGATACTACAGTTTTTTGTTCTCAACAGGCCGAGCGACAAGAAGAGATAG | CTATATATCTATTAAGTTAGGATTTTCTTGCTGATGCTGCTGTTGGATCCCAATACAACAGAT |
YIR013C | CTGTATTAGCTTGTCATCTAAGCCTTCGTCATTGAAGTATCCC | AGCGACCCTGTAATGTTATGTTTCTAGCTAGGAACAGAAAGTG | AGAAACATAACATTACAGGGTCGCTCCCATTATACTTTTTCAGCTTTCAACCTATCGGGACAAAACACACATTACTTGCCTC | TCAGAATCTGACAGTTCCGGTAAGTTCTTTGGGGATCCTTCTCTTGATCCCAATACAACAGAT |
YJL065C | TGACAGTGGGTATTGCTAGGAAACTCTCTAGGGACAAGGCGCAT | TGTGTTAGCCTGATCACATCGAGCGAGCTATCCTGACGCTATTC | CGCTCGATGTGATCAGGCTAACACACTACACAAGCACATTGCATAGAAGGATTTTTCTTCTCTCTCACACATTACTTGCCTC | TTATTCTTGATCAGTAACGTCTTCGTCGTCGTCTTCGTCATTATCGATCCCAATACAACAGAT |
YJR153W | AGGGGTTAAAGGGCATTGAGTTTTTGTGAGTGGAAATGCCCCCGTTATA | TGCGTTTGTCCATCAATGTGGGTAGATAGTCTTCCCGTTTGAAGTTCTG | CTACCCACATTGATGGACAAACGCATCCTCTTTTAAAGTACTCATATGACTATACATACCTTCTTCACACATTACTTGCCTC | TTAACAGCTTGCACCAGATCCAGATGGGATTCCAGAACATCCAGAGATCCCAATACAACAGAT |
YKL132C | GAACGAGGACCTAACGCTAACAGCAGAGTCCATAT | CTGCTTCTTATGTAATCTGTTGCCGGCCAAACGAG | CGGCAACAGATTACATAAGAAGCAGTTTTTAGATATATGATCATATCCTACAATGCAATTACCGACACACATTACTTGCCTC | CTATACTGGCAAGTGACAGTTGTGTAAGCGCAATAGCTCTTTGCAGATCCCAATACAACAGAT |
YKL191W | ATATAATCGACCGTTTCACCAACACTTCTCAACTCATCAGCCACTTTGTC | TTTTCAACCTTTGCACGATCTTTCAACGCTGCCACTTAGAAATCCATCAC | TCTAAGTGGCAGCGTTGAAAGATCGTGCAAAGGTTGAAAACTCTTATAATTTGTTTCCCTCGACTTCTCTATTTAAATCCCACACATTACTTGCCTC | TCATTTGTTTTCCTTTTTCATAGCGTCTTCGCGATCAAATCCATAGATCCCAATACAACAGAT |
YLR065C | CCACGGTCAGTGTATAGTTGCCTACAGTTTCACCGTAAGTTAGCAAACCTACCCA | TCTACCAACACAAACTTAGACCACTGCCTTTGGTGTCTTCTTGAAGCCACACGTG | AGTGGTCTAAGTTTGTGTTGGTAGAACAAAGCTTTTCAAACGTTTTTGAAATTTTTTGCTTTCTACACACATTACTTGCCTC | TTAACGATACTTGTACTTAATTTTAGAGTCAGTTTCACCCCTCCTGATCCCAATACAACAGAT |
YLR079W | TCATGATGGTCTCTGATGTACTAAAGGGGTTATCGGAGTCCATAACATGCGAT | TTTCGTGTAATAGTCCCTGCTTCAAGGGTCAAAATTCCGTAGAGGTTTGGTTG | TTGAAGCAGGGACTATTACACGAAAAGAAATGAAAAAAAAAAGTTTCTGGAACGATTATATTTTAAGCGACAAGAAGAGATAG | TCAATGCTCTTGATCCCTAGATTGAAACAATGCCTTTGGCTTGAAGATCCCAATACAACAGAT |
YLR091W | AGCTTCGCAATGGGTGGTCTGTGAGAAATG | TGTTGCTCCAAGTTCGCAATCGCTGTTCTG | CAGCGATTGCGAACTTGGAGCAACATTGTTGTTGGCTGTTATACTGGAGTAAATGTCACGTGCCACACACATTACTTGCCTC | CTAATCAACTTCCCACCGGGTGCAATGAAACATCAAAGGCTCCGTGATCCCAATACAACAGAT |
YML097C | CCTGGGATGTATGGACTCTACCCTTTTTTTTACAGAATCACGTAGCACAGCAT | GGCTGTTGATTCCTGTTCTTTGTTGTTTGATGAAGAGAAACTCAAGCCCGTTG | CAACAAAGAACAGGAATCAACAGCCTGATCTCATGCACATATTTCATGTATAGAGTAGCCTAGCACACACATTACTTGCCTC | TTATTCTGACAGAGAAAGTAGAGCATCAACACAGGGCCCAATACGGATCCCAATACAACAGAT |
YMR035W | TAGCAGCCTTTACCGCGCAACAAAATCCATATACTATGGACGAG | TACTGTACAGTACAATGCAACAGCTTCATGAATAGCAGCACGCC | AGCTGTTGCATTGTACTGTACAGTAAAATCAGCTTATTTTTATGAATTTAGAAGTGACCCCGGTACACACATTACTTGCCTC | TCATTCTAAAATGGCTCTTTTGCTAATGCAGTCTCTACCGGTACTGATCCCAATACAACAGAT |
YMR103C | GATAATGACAAAACAGACACTTTCTGTGGTACTCC | TCTATTTTCTGTAGTTCTTCTTGTCCCCGTTAACC | ATTGTGGTTAACGGGGACAAGAAGAACTACAGAAAATAGATCCCGTAAAGGGAAAAAGCTAACCTCATTCGCCTCACAGGAGCGACAAGAAGAGATAG | CTAGGTAAGGCTCCCCATGGCTGAGCCCGCGCAAGGAAATCCAGGGATCCCAATACAACAGAT |
YNL129W | ACTTTCTCCAAAAGGGCCTCAATACATGGTT | ATTTTGAGGTTCTACTCTAGCTCACACTTCG | GTGAGCTAGAGTAGAACCTCAAAATGAAAGCGCCACAAAATCGATGAGAAGTATAAAAAAAAAAACACACATTACTTGCCTC | CTAATCCTTACAAAGCTTTAGAATCTCTTGGCACACCCAGCTTAAGATCCCAATACAACAGAT |
YNR057C | TTGAGATCGGTAACTTCTTCTGCATTCCAATGGTCCTCGAAACCGTT | GACCTGTGCGCTAGTGATTTTTTGAGTTCTTTTCACTGGCCTACTTG | TCAAAAAATCACTAGCGCACAGGTCTTTAAATAAAAAAGTTGCAAGGCTTTCATCAGCTATATATAGCGACAAGAAGAGATAG | CTATTCTACGTCTTGTTGGGTCATTAATGACTCCACCGATGGCAGGATCCCAATACAACAGAT |
YOR076C | TTCAAAAGCTCGGCGGCTATGACTTCTTGCTCCTCTTGTTGACTTCGCA | GTCAGGTGGTACTGTAAGTTTCGAAGACCACCTCCTCGTACGTGTATTG | TTCGAAACTTACAGTACCACCTGACTCCTACAACTAAGAAATTATACTAGGCATTCATACTTATTCACACATTACTTGCCTC | TTACTGGCATGCAATTCTGCCAACTGCGATAACCTTATGATCTATGATCCCAATACAACAGAT |
YOR106W | GTAGCAACAACGCTCATGTTGCTTCACAATCTCCAATTTTTGACATCAGCGATTA | AATCTCAACTTTCTGCAGTGGATATTAGTTGGCCAATTTGTTAATGCTGATCCCG | CAAATTGGCCAACTAATATCCACTGCAGAAAGTTGAGATTTAATCTCCTTAAACGCGCATTGAGCACAGACTTTCTGGTACACACATTACTTGCCTC | CTAACTTAATACAGCAAGCAATACCACCATGCACACAACTATTATGATCCCAATACAACAGAT |
YPL202C | CCCATAGAGAATTCAAATTCTTCTCCACCCGTCTC | TTCTTGGGGGTCGCTTTCTCTTGATTAATTTGCGC | ATCAAGAGAAAGCGACCCCCAAGAATTATTTAGTTTTCAACTCATTAAAACGGTATCACGTTTAACACACATTACTTGCCTC | TTAATATTTTGATATTAAGGCTGGGTCAATAGAATTTAATTCATTGATCCCAATACAACAGAT |
YPR054W | AGAGGAAAAGAAATTAAATCACACAGACAAAAACGCGGGGAGGAAG | TCTTGGAGCGTTGAGCACGTCTACATTTCTATCGATATGATGTATC | TGTAGACGTGCTCAACGCTCCAAGATAACCACTACACAAATAGGTATATCATGAAGTTATATTCTCACACATTACTTGCCTC | CTATAAAGACGAGGAGGACAAATCGGTTTTGAAATTTTTCACTTCGATCCCAATACAACAGAT |
YAL058W | CAAGTACATTCAAAATACAAGAAAGGCTGGCTGGGATAGCACCAATGAGACTATAAAAA | AGTACTACTTTGTGTGGCCGTTATCTATATTAGAAATGCGTTGCCGTTCACTTTTTTAC | GATAACGGCCACACAAAGTAGTACTTGGACAAGTGTTAGTTTATAACATGGTCTCAATAATTGCACACACATTACTTGCCTC | CTATGTAAATACTACACAACAAAGAACCGACGTCGTCAAGAGCACGATCCCAATACAACAGAT |
YAR028W | GTTGTAAACCGCTAATGATTATGTTCCTAATACCTGTGTGTTTGGTTT | GGTCAATTGATATCTTGCCTTAATATGGCACTTATAGATGTTATCCTG | CATCTATAAGTGCCATATTAAGGCAAGATATCAATTGACCAGCGGAAGCATTTTATTCACCAAGTATACTTACTTTTCTTACACTCCTCAGAAGCTC | TCAAGGTAAATCAGCGTCAGGATATTGCTTTCTCCAGTATTCACTGATCCCAATACAACAGAT |
YBL033C | GGTTCAAAAAGTGGAAGTAGTAGTGCTCTT | CCTGTAGGTTTGTTGTAGTTTTATGTACGC | ACAAAAAGTGGCGTACATAAAACTACAACAAACCTACAGGACACAAGTGTAGCATAATTATTAGTACATATAGTGCTTTTAGCGACAAGAAGAGATAG | TTATATTGCCAGCGTCGATGTTGATGACAACGCAGAGTTCATACGGATCCCAATACAACAGAT |
YBL055C | GAGAAAGTGAGTGGGAATGGACAAGATTATTTGGGTGAGGGTGTTTGCTTA | ATAACTATAACAGTACTGGCGCCTCAAAAAAATGCCTTGGCAGCGAAAGTG | GAGGCGCCAGTACTGTTATAGTTATCATCTATTGGTGCAAAGAGCAAAGTTACATTATTTTTAGTAGCGACAAGAAGAGATAG | TTACTCTCCAAATATCTTACAGGTCGTCTTCCACGTGGTATCTATGATCCCAATACAACAGAT |
YBL091C | TGCTTACTTTTCGTTTCCACTTTATCTTGCTGTTCGTTCGAGCAACAGTATTTATT | TTTTCAATACGGTAGAGCTTCTACAGTACTTGTTGATGTAAAAGCCCACTTATTAG | TACATCAACAAGTACTGTAGAAGCTCTACCGTATTGAAAAGGTAAAATGCGCTTTCAAATGGCCTCCTCACTAGGTATATCACACATTACTTGCCTC | TCAGTAGTCATCACCTTTCGAAACGACTTCCTTTTTGTGAGCATGGATCCCAATACAACAGAT |
YBR077C | CGTAGGTGTTTGACATTGAATATTTATACCGGCT | ATTGCTTCAATATTTTGGATACCGTTTGCTGCTG | AGTTATCAGCAGCAAACGGTATCCAAAATATTGAAGCAATTGGGTTTAACAATATGTATCGAGGCCACACCTTTTCATGCACAATCATATGGGAGAAG | TCAACCTAGCTTGTAGCCAAACAAATCAGTCAACTCTCTCATAGCGATCCCAATACAACAGAT |
YCL050C | CTTTTTTTTATTGTGTTGGTTTTTATATGTTTTGTTATGTATTGTTTATTTTCCCT | TTTTATTTTAATTTTTACTTTTCTGTTTGTTCTAAAATCTATCTAAACTGGCTTTC | AGATTTTAGAACAAACAGAAAAGTAAAAATTAAAATAAAAGTCCTTCACAAGTACAAGTATACACGTATACATGTATGTACACACATTACTTGCCTC | TCAATAGTTGTATTCGTTTGGTTTTTGACCAGAAGTGTTTGGGAAGATCCCAATACAACAGAT |
YDR071C | TTTGTTTTACAACCAAAAGTTTTAAAAAAGTGGATTCATATAAGGTTAG | TTTAACTTTTTATTTAATCGTAATGTATTGGTGTTATTTTTTCTCCGAG | AAAATAACACCAATACATTACGATTAAATAAAAAGTTAAAATCATTAGTTTAACGCACTTTACAAATGTCTAATTATAAACACACATTACTTGCCTC | CTAGTTGTCGTATTCTTCCTTAATTAGTTCTCTCTCCATATCGATGATCCCAATACAACAGAT |
YDR330W | CACGATGTAACCCAGTTATAGAACCTATTCTGTAGTTTGTTGCAGTTTT | ATAAAAGGTATGTTATTATTGCCACACGTCGTAGTCTATGCGTTTGTAG | CATAGACTACGACGTGTGGCAATAATAACATACCTTTTATACAAATTGTATGGTGGCAGGTACCTAGATGTAATTAGGTAAGCGACAAGAAGAGATAG | TTATTCTATTTCAGGGTCAAGCTTCTCAAGTAGTAAAGAACTATTGATCCCAATACAACAGAT |
YER115C | AATAGTAAGGAACAAACTGTTGATTTAACTGCAGATGCCATAGACTTA | ATACAAGTTTGAACAAAATACAACTCAAAGTAAAATGGTAGCTGCACG | TACCATTTTACTTTGAGTTGTATTTTGTTCAAACTTGTATACACAGGAGAAACAATTCCATAAATAAACTTTGTAAGATACACACATTACTTGCCTC | TTAAAAATATAGGGATGAAAACTCATCGGCTATACTGTCATAGACGATCCCAATACAACAGAT |
YHL044W | GCAGTACTACAACTCTGTGCGGTGGGAAAAATTTATTCTTCGAGGCACAGTATT | AGCCTTAGCGGGCCCTCACTTGCTTTTGAATTTGTCTTTATACCACAAGTTAAC | AAGCAAGTGAGGGCCCGCTAAGGCTTTGGAAGGTATTTCTAGGTTCTACTTTAGTTTTTTATAACCACACATTACTTGCCTC | TCAAACTGTGACCCCCATCTCTGGATACTTATCCTCCCAGTACTTGATCCCAATACAACAGAT |
YHR207C | AAGGATAATAACAGAATTTAATGGCTTGATATGGACGGGCG | TTTTGTGCCACTTTTCGTTATTTTGTATCAGTGCACGCTTG | AAGCGTGCACTGATACAAAATAACGAAAAGTGGCACAAAAGAGATAAACGCATACTAAATTACTGTGTCTGTTATTGTTTCACACATTACTTGCCTC | TTATCTTTCATCCACTGCGACCGAAACGTTTGAATCGAATCTGACGATCCCAATACAACAGAT |
YIL079C | CTTTCCTCGATGAAAGTTGGTTCAGGATCCAAGTACAAGGACTGTTGCTT | AATATCTTAGCTCCGTGGTCCGCTTCCTTTGTATTCCTCATGTGAGAATG | AAGCGGACCACGGAGCTAAGATATTGATTATATGTGAGTTTTTTTAATTTTTTTTCCATTCTCATAGCGACAAGAAGAGATAG | TCAATAACGGCCATTTCTTTGGCTTTTGTTAGAAGATGATCGGCCGATCCCAATACAACAGAT |
YIL111W | ATGATATTGTTGAAGGATATCGACTTGCTCTCAAAGG | GCTCTTTACAATGAATATGTGGCGTTATTCCATTGTG | ACGCCACATATTCATTGTAAAGAGCTTTTTGTTTTGAGTTTTAAAAGATATCCATTTACTAACATCACACATTACTTGCCTC | TCAGTGTTTGTTATACATGCTCTTTACAATGAATATGTGGCGTTAGATCCCAATACAACAGAT |
YJL058C | CGGAAGTGAATGAGAATACCAACAGTTTTACTGATGATTATATTGACGGAG | AATGAAAGAACTTTATTTCTACAGAGGTCAATTGGCGTTTCTGTCACCTAG | AAACGCCAATTGACCTCTGTAGAAATAAAGTTCTTTCATTAAAGCATCTAACACTTTATTTCAATCTCCTATTTAATATCCACACATTACTTGCCTC | CTATATACTCATTCTAATGCCTGCTAATAACTCTTCGATAATTCGGATCCCAATACAACAGAT |
YJL128C | GTAGTATAGTATTGCAACAGGAAAAAAAAATCTTATTGCGTAGCACTCTGTATCTATTTT | CTTTGAGAAATTAACGTCTCGATCTGCTTAATATAATAATGTATCTATAGAAATGTACGG | ATTATTATATTAAGCAGATCGAGACGTTAATTTCTCAAAGCGTTAATATCCAAATAAAAGCAAACAGGCACGTGAATATACACACATTACTTGCCTC | CTATAAACCACCCATATGTAATGCCGGTACATTTTTAGATAAACCGATCCCAATACAACAGAT |
YKL023W | TTCCCCATATTGGGAACCTGCTCTGCCCTGCTCAGCACGTAG | GCTTGCACGTTATACCTGGCCGGGATTGGGGCGCTATTCTTG | TCCCGGCCAGGTATAACGTGCAAGCATGCATTCTAATGACATAACTATTTGACGACGTGAATTAGAGCGACAAGAAGAGATAG | TCAAAATTCTATAGGCTGTCCATTCATGTCAGTTGAGGAAGGAAAGATCCCAATACAACAGAT |
YKL166C | CCAGGTACGAGTGATTTAGGAAGGTCTGAAGAAAAGCGACTATGCATT | TTTGTGCAGGCTCGCTCTTTCCTTGTACAACCACCGATATACAATATG | CAAGGAAAGAGCGAGCCTGCACAAAGGAGGTTCAGTGGGATAATTGTAATTGAAAAAAAAAAAAACACACATTACTTGCCTC | TTAAAATTCTTTCATTAAATCCATATATGGATCCTCCCCTTGAATGATCCCAATACAACAGAT |
YKR039W | TTATTTATTTTTTTTTATACCATTTCCTTTTGATAAGGGGT | TTTTTATTTCTTTTTTTTGTTTCTTATAAATGTTGCTGTCC | GACAGCAACATTTATAAGAAACAAAAAAAAGAAATAAAAATGAATCGAACAACGACAAAAAAAGACTTTATTTTTTAGATCACACATTACTTGCCTC | TTAACACCAGAAATTCCAGATTCTATACCATCTTGGCTTTGTGGCGATCCCAATACAACAGAT |
YLR218C | GTAGTCTTTACGGTAATGGAGTTGGCCCTTG | AAATGAAGCGAATGGATGAGAAGAGGGGAGG | CTCTTCTCATCCATTCGCTTCATTTAAATCTAAATGTCGTGATGTATAACTTGTATATAATAGACCACACATTACTTGCCTC | TCATTTCTTCTTTTCCGAATCCTTACTGGTCGTCCCATCCACGTCGATCCCAATACAACAGAT |
YLR255C | AGGGTGGAGAGTTAGAGCTGGTGTTGGCACCGGGCGTAGTTGCTG | TCACCCGTCGGGCGAAATCCACCCTCGCGGCCTCGTTTAGAATAC | AGGGTGGATTTCGCCCGACGGGTGATCAATATTTCTTGAAGTACGATTTCTGTGTGTATAGATTCCACACATTACTTGCCTC | TTAGCGAGCCTTTAAGCACAGAACAATCTCTATAGCCAACATCTTGATCCCAATACAACAGAT |
YLR350W | ACGATTATGTATATCCATATATATATATACACTACAAGCCGACCG | GATGAAACTGTTTCTTATGCGTTTTTATAATGGTATAGTCTTGCG | GACTATACCATTATAAAAACGCATAAGAAACAGTTTCATCTTGGCTCTGCCTATACGCATATATGTATATATATATATATCACACATTACTTGCCTC | CTAACTAATTTGAGCACGGCCCGTAATACCAGGGATGGATATTCTGATCCCAATACAACAGAT |
YML048W | GCTTTTGCCCTTGACCCTGAATGCGGCAGATCCAGCGACTTCATCGAGGTTA | GCTTGCAATCGCCCTCTATCAAAACAAAGTCCACCGATCTCTGCTGCCCCTC | GTTTTGATAGAGGGCGATTGCAAGCTTGTAAAACCAAAACACTACCATCCAGACTTTTTTTTTATCACACATTACTTGCCTC | TTAATTAGATTTTTTCAAATCATTCTTTTTTATACCAAAAGTAGCGATCCCAATACAACAGAT |
YML088W | CTCTGAAGAGTTTAGCGGGCTCTCAGACGGTTTTTTCGCTTTTGGCATTTT | GCTGTCTTATTCCCTAGTGTCGGCCACCTGATAAAGGTTTTTAACTCGTAC | GGCCGACACTAGGGAATAAGACAGCAAGTATGTACATTTGGAAGAGCATATGTTAAATATTTTATCACACATTACTTGCCTC | TCAATTGATTTCACTCAATGACAACGCAATGGCCAATTGTAAGTCGATCCCAATACAACAGAT |
YMR018W | AATTATATTCTGCGCTTGCCTTTTATATTAAGCGTCACAACGCC | CGTGCTTACAAAAGGTACCCTTTATTCTTAATGATTCTCCACTC | ATAAAGGGTACCTTTTGTAAGCACGGTTTTAGTTAATTTTTTTGCTATATAGTGCATTCAAAATACACACATTACTTGCCTC | CTAATCCATTTTGTTATAAATAGCTTTTAAGGTTGATATTAACGAGATCCCAATACAACAGAT |
YMR278W | TTTAATCACCAAGAACCCTAGCACTTATAAAGCGCGTCTAGCGAAAAAGTT | TTTTTGAAATTGCAGCAGCTAATGGCTCGAGAGTACTGTTGAGTATTTAGG | AACAGTACTCTCGAGCCATTAGCTGCTGCAATTTCAAAAATACTATTTTAACCTATACATTAAGACCAAGCGACTTAAAGCACACATTACTTGCCTC | TCAAAATTTTGTAACTATATTCATTTCATCTGGTCTAAACCATTCGATCCCAATACAACAGAT |
YNR058W | TAGTACATTGTGCAACCTGTGCCATAATATTGACACCGAACTTCTTCAGGGCCT | TAGTTGTTATCCTAGCAATAGAAAACGAATTGCGCTGTTGACATCCTCCCGACC | CAACAGCGCAATTCGTTTTCTATTGCTAGGATAACAACTATATACGATCTCATCAAGTCATTTGCCAAGGAATTCTTATTCACACATTACTTGCCTC | TCATTGGTTTATGTGGGATTTCCACTCATGTAATACTTCAATCAAGATCCCAATACAACAGAT |
YOL158C | AGAATCATCTCTACTACACACTGCATTATGAACTGCAGCCACATTTATCTAAGGG | TTTCAGAGAGACATTCACGAGCACTTTTTTGCTATATTAGAGTTCTGCTAGGACC | AGTGCTCGTGAATGTCTCTCTGAAAGCTGGAGAGTACTAGCGATACAAATATAGTGTTAAGCTCTAGCGACAAGAAGAGATAG | TCAATATCCAATTACACGACGGAGCCATGATTGTTGTTTGATTTTGATCCCAATACAACAGAT |
YOR008C | AAATAATGTGGAGATCTTATCTTGTTTTTCTCTTCTTTATGACA | TATTTAAATAGAATTTTTTATCCTGAGTACTAAATCAGCCAACG | GGCTGATTTAGTACTCAGGATAAAAAATTCTATTTAAATAAGAAACCCAAAAAAAATTATTATATTCTTAAACTATTGCCCACACATTACTTGCCTC | TCAATCAGCTTCGTCTGGATTGACCACTGTTAAAACGTTGTTTTTGATCCCAATACAACAGAT |
YPR037C | TGCCCTTATACCGCGTAGTGTAGGTCCAGTCGTACTT | CCCTTCGTTCCTTGCACAATCTGTCCAGACTTTCCTC | GACAGATTGTGCAAGGAACGAAGGGACAAAGAAAGATTACATTAATCAATTATAGTAATAGTAAAAGCGACAAGAAGAGATAG | TCAACCGTGCTGTTTAGCCTCCTTCTCGAGAGACACGCGTTTACCGATCCCAATACAACAGAT |
YPR040W | GCCGTCCTGCTAGATGTCTGTACGGGATACTTCTCAATCAACTT | TTTTTCTGGGGAGCTGTTGTGTCTAAAGCTGCCCTTAGGTCATG | TAGACACAACAGCTCCCCAGAAAAACAATCAAATATACAAAACAATACGTACAAATACATACACGCACACATTACTTGCCTC | TTAGAACTGAATTATTTCACATTGTCTTTTGATCATCGGCGTGTTGATCCCAATACAACAGAT |
YPR051W | CAACCAAGGTTCGTCGTCAGTAAGCTAGTTACGGAATGGTAA | GTCCTGACCCAACGCCTTTTCTGTAGTAGTGGCTTTTTTATC | TACAGAAAAGGCGTTGGGTCAGGACAAGGAAAAACACTCGATGATGATAAAATATATATTAATAAAGCGACAAGAAGAGATAG | TTATGTGGCCAGCCGGCCATGCATTAGGAACGTAGACCGAGTGCAGATCCCAATACAACAGAT |
YBR273C | TAGGTTCAAGTGAGTAAATTCTATCTTTGACCAGGGTCCCCAGTTT | TTGGTTACGGCTGTTTGGTTTAGCTATCCTTGCTTCTTTGTAAGTG | AGCTAAACCAAACAGCCGTAACCAAGTAGTTAATGGCTTCAACTCCATTTATGACTAGTATTAGACACACATTACTTGCCTC | TTATTTTTTATCTTCGTCCTTTTTCTTTTCTAAATGGACATTATTGATCCCAATACAACAGAT |
YAL030W | ATGCACGATTTATAGTGATCTAGGGGTACCTCCTCGG | GGACAATGAATACGTTGCGCTTGCTATGTCTTTGGCG | AGCAAGCGCAACGTATTCATTGTCCCTTTCTACATCGTTGATACTTATGCGTATACATTCATATACACACATTACTTGCCTC | TTAGTATAATGTTAATTTCCTAGATGGTACTCTGTGGCTTTCTGTGATCCCAATACAACAGAT |
YAL065C | CATTCCAACCAGTAGTTCCACCTCTGGTTCTTCTGAGAGCGAAACTA | TTTAGAGCTGACAGACTCTTCATTCGTAGTCTGTGGCCTCCATGTTG | GAATGAAGAGTCTGTCAGCTCTAAAACGCACGGCATCATCTTATATTAATAATTCTATCATCACGCACACATTACTTGCCTC | TTACCATACGATTGCCAGCAATACGGTGGAAATAAAAACACTTATGATCCCAATACAACAGAT |
YAR002W | CTCGATTTGCATAAGTGTGCCTTAGTATGC | TGGATGTCGATTTTAGGATATCTTGCGGTG | ATCAAATAAGCACCGCAAGATATCCTAAAATCGACATCCATAATGAAAATTTTAGCCGTGACATAATTACCGTATAGCCCCACACATTACTTGCCTC | TCAAAAGGTATATAGGGACTTGAAAGCCTCAACTTTATTTTCATCGATCCCAATACAACAGAT |
YBL049W | CGTGCACCTTTTCCTCGAGATAAACATATAGCGATGACTTAGAGAACAA | TTTCTTCTACAGTAAGATAAGCTTCTCCCTTTTCCTTCAGTGCTGCTAC | CTGAAGGAAAAGGGAGAAGCTTATCTTACTGTAGAAGAAATGTCTTCCTTTGTCTGCTATCTAGCACCTCTCGTCTTTTACACACATTACTTGCCTC | TCAAGTACATTTACAAATGTTTTTCAGCTCTAAGATAAACTTTCCGATCCCAATACAACAGAT |
YBR144C | TGCTTCAGAAACAGACACATTAATAACTCCATGAGAACCGCC | GTGACAATGAAATAATCAAATTGTGACATCTGCTGACGCGGG | CGCGTCAGCAGATGTCACAATTTGATTATTTCATTGTCACTAAGAATTAGCAACTACTTTCATTCATCTTCGACGCAACTAGCGACAAGAAGAGATAG | CTAGCACATGAAAAAAAAGGATATATATAATTTTACAAGTAGTGTGATCCCAATACAACAGAT |
YBR185C | ACATTTGATAACCCTCAGAGATGATCCTCG | CTCATAAGCTTAGCTGAATGGATAGGCTTG | CTATCCATTCAGCTAAGCTTATGAGGGCAATATGTATATAAATGCGAAAAGTTTTGGAACATCTGAGCGACAAGAAGAGATAG | TTAGCTTGGAGGTAAACGATATATATCACCGTTAACTTTCATTCTGATCCCAATACAACAGAT |
YBR255W | GCTTACGTGCTAATGTCCTTACCCTCGAGT | GTTTCATCCGTTTAAAGTACCTCCCCCTCC | GGGAGGTACTTTAAACGGATGAAACTTTGCGGGTCTATGTATTTATAGAAATAAACCATCTGCACCACACATTACTTGCCTC | TTACCACAGCAATGCCTTAATGATTTTTAGAACTAAAAAAATTGTGATCCCAATACAACAGAT |
YCL044C | AAAACGATGTCAGAGGCGTCGTAATTGAAGGTTACCCAACAA | CTTTTATTACGGTAATTGGAAAAGGAGAGGGAATGGAGGAGG | TCCTCCATTCCCTCTCCTTTTCCAATTACCGTAATAAAAGGTCTTTGCGGATTGCTTAGTTGTACCGTGCGTATATTAAAAGCGACAAGAAGAGATAG | CTAATGTGTCTTTTCATTAGTGAGAGCCTTGGGGGTAGGCCCTGGGATCCCAATACAACAGAT |
YCL047C | GAGAAATCCTCATTACCCAAGAGCACAAGGAT | ATTAGTCTGCTAGTGCACGTACTGCTATCAAC | GCAGTACGTGCACTAGCAGACTAATAAGAGAAAGATAGAAATTCAAATGCCTTTCATGGAGATATCACACATTACTTGCCTC | TCAATCAAATATTGTCTTATTATTGATCAAATAATCGTATATGCAGATCCCAATACAACAGAT |
YBR268W | CACAATAAGCACTTCAAAGATGTTGAAGAAAACCTGGAACACATGTTTAGGACCC | CTTCAGTCTTCAGCAGCTATTTCCTTTGCTGTAGATATAACTGCTTTAAGAAGCC | AGGAAATAGCTGCTGAAGACTGAAGGCCTCCGTTAATAAGATCAAGTTTGGTATCCTTGTAAATACACACATTACTTGCCTC | TTAAAGTTGGCTCAAGAAATTGTTTTGCTTGATTCGCTGCCTGTTGATCCCAATACAACAGAT |
YAL002W | AAGCCAGCTGCTAAGTCCATTGTCACTCTA | TCTAGGTGTAATGAGTAATGGTCTCGGCAG | GAGACCATTACTCATTACACCTAGACATTTCTAAATATTTAATACAACTTTGGTTACATAAAAGTCACACATTACTTGCCTC | TTATACTATTTTTGGGTTAGATTCCGTCTGGCAAATTAAACAAGAGATCCCAATACAACAGAT |
YBL082C | AGTGTTGTTGTATTTCATGTGATTGTAATGGGGGTGGTAG | ACTACTAGTATCTAACTCTTTCTCTCTCCCATACACACTC | GAGTGTGTATGGGAGAGAGAAAGAGTTAGATACTAGTAGTAACAGATGTATAAATATCATATAATCGTATTGTTTTAAATCACACATTACTTGCCTC | TCAGTTGAGCTTTTTTTCCATAGAGCTGGTGGTACGAAGATGGCTGATCCCAATACAACAGAT |
YCR045C | AAGAAGAAGAAGAAGAATCGAGTCTTCCCTCTGAG | GACCAGTGTTTTTTTGCTGTTTTTCTTGGCGGATG | TCTAACATCCGCCAAGAAAAACAGCAAAAAAACACTGGTCTGTTTTTCTCTTTCTCTCATAAATAAATTATTGCATAATGCACACATTACTTGCCTC | CTACAAATTTTTCATAAAATCACGCCTCTTCAATCGAATCTCACCGATCCCAATACAACAGAT |
YDL106C | CTTCTCGACTTCTTTTGTTTTCTTGTTTCTCCGCT | AGCGTATTGTATACGTTATTTAGCAGTAAGCCGTG | CAAGTCACGGCTTACTGCTAAATAACGTATACAATACGCTTGAAAATAATTCTGTTTTTTTTTTTTTGCGATTGCATTTTCACACATTACTTGCCTC | TCATATCCATCTATGCTCGTCAGTTAGTTCGTTAGTGTTCTTCAAGATCCCAATACAACAGAT |
YIL148W | GTGCATCTATTTCCCGCCTATATCTGGAACGTTCTTCATT | GTTTGAATCTTGGTCGTCTCAACTATGGGATAATAAGCGG | TAGTTGAGACGACCAAGATTCAAACACATAGTTTTTCGAACGTTCAGAGCTTAAAGGGACAATTACACACATTACTTGCCTC | TTAGAATGGATATGGTGCATACATTTGCATGTTTGAATCTTGGTCGATCCCAATACAACAGAT |
YJR073C | CGTGAAACTTCCGTACCATTTCTTGCAACAATTGGTAAACAGCA | CGCTTGACTTGCGCTATTCTTGTTGTCTTCAATTGCTGTTTATG | CAACAAGAATAGCGCAAGTCAAGCGGTGCTCTATTTCTATATATTCTATAATGTTAGAAGCCTATCACACATTACTTGCCTC | TTACATATTCTTTTTGGCCTTATCACGGTTAGCGTAGATCATGGCGATCCCAATACAACAGAT |
YLR011W | CGCCGAGCTTGGCCATTACAATATAGAGCACATGCTATGA | GTTTAATGCGGTTGTCGACTAAGCACCTCAGCTGGTAATG | TGCTTAGTCGACAACCGCATTAAACACCATAAAATTTAACTGTAGTCGCAATAAAAGATGAAAAGAGCGACAAGAAGAGATAG | TTATTTATTCCTCGTTGTTTCGATGCACGATGCGAGTAATTGCAGGATCCCAATACAACAGAT |
YCL044C_2 | TGTTTTGTACTATGCCCCATGGTGTGGTCACTGTAAGAGATT | CTTTTATTACGGTAATTGGAAAAGGAGAGGGAATGGAGGAGG | TCCTCCATTCCCTCTCCTTTTCCAATTACCGTAATAAAAGGTCTTTGCGGATTGCTTAGTTGTACCGTGCGTATATTAAAAGCGACAAGAAGAGATAG | CTAATGTGTCTTTTCATTAGTGAGAGCCTTGGGGGTAGGCCCTGGGATCCCAATACAACAGAT |
YDL022W | TATTTTGTACAGCTGATGGGACCTTGCCGTCTTCATTTT | CTTTATATTATCAATATTTGTGTTTGTGGAGGGGGGGGG | ACCCCCCCCCTCCACAAACACAAATATTGATAATATAAAGATTTATTGGAGAAAGATAACATATCATACTTTCCCCCACTCACACATTACTTGCCTC | CTAATCTTCATGTAGATCTAATTCTTCAATCATGTCCGGCAGGTTGATCCCAATACAACAGAT |
YDL175C | CGTCTACAGAACCCCTGAAGAACACGAAACCTGTAGCCAA | GACTGCGAGGAAGTGATGTAAGCTGCTATCGTTGTACAGG | CAGCTTACATCACTTCCTCGCAGTCCTTTAAATAGGAAGCCCTTTAACAAGCTGTTCTTGGTTACCACACATTACTTGCCTC | TTATCTTCTTTTCCCCAAGGTACCACTCCTATATGGTTGATAAGAGATCCCAATACAACAGAT |
YGL211W | AAACTGCTGACGAAGTGGGATGGGATGAGATGATAA | GACGGTTCCTTTGCTTTACTCTTACCATATCGTCTC | GTAAGAGTAAAGCAAAGGAACCGTCCATATATATAGCTCGCAGTAGAAGAATGCAGCGTAACATAAGCGACAAGAAGAGATAG | TCAAAAGCTGAGTTTCTCAAGAGCTCTCAGCTTCAAGGCTGCACCGATCCCAATACAACAGAT |
YAL009W | TTGTCACTACTCAACGTATTCGCACTACTAACACTG | GACCTCCTTTATTTAGCTTTCCACTACCTTTCTTCC | AGTGGAAAGCTAAATAAAGGAGGTCAAGAGTTGGAGGGCTTCTTCCTTCGAATAAGAGGTCATATCACACATTACTTGCCTC | TCATTCTGATTTAGGTCGGAGTTCGTGCGCTTGTTTGCGTCTTCTGATCCCAATACAACAGAT |
YBR185C_2 | AATACTTCCTTGCCCCTCTCTCATTAATTCTTGGCCAGTTATAATCGA | CTCATAAGCTTAGCTGAATGGATAGGCTTGCTTTCTGATGGAAATTTG | CTATCCATTCAGCTAAGCTTATGAGGGCAATATGTATATAAATGCGAAAAGTTTTGGAACATCTGCACACATTACTTGCCTC | TTAGCTTGGAGGTAAACGATATATATCACCGTTAACTTTCATTCTGATCCCAATACAACAGAT |
YBR270C | AATATTTCGTCACGTTGCGGTGAGCAAGGATCTTTTGGCT | TGCTTGACTCTGTAGTTTAAGACTCGCCTAGCCACTTATG | GAGTCTTAAACTACAGAGTCAAGCATTAAATATTTTATTTAGTAATATACAATAGTGGCATATAGCACACATTACTTGCCTC | TTATATACTCATGCGTATGCCCTCAAGAAGTTCACCAATAATTCTGATCCCAATACAACAGAT |
YCR025C | TTATTGGTATGGGGCCATATTTTCCACAGGGATACATTGAGC | AAACCTCCTCGTTTAGGAAAGAGTTAACACGCCTAGTTTCAG | AACTCTTTCCTAAACGAGGAGGTTTTAGAAGATAGCTTCAATTTTTTCCGATAATAATTTCCCATCACACATTACTTGCCTC | TTACGCAGTGACAAGTCTATATTGACATCTTTCTGCTTTCTGCTTGATCCCAATACAACAGAT |
YDL012C | CGCTAGGTTGCTAAGGTTTCGTCTTTTCTATCTTACGAT | GAGATGTGTATTTGCTTGGTTTATGGGCTATTGTATGGG | ACCCATACAATAGCCCATAAACCAAGCAAATACACATCTCTTTTACAAGTTTTGCCTTTAGAAGATCACATCGTACTAACCACACATTACTTGCCTC | TTAACGTACGCTTTGTTTGGAGGCAGAGTTTCCGTAATAATCTTGGATCCCAATACAACAGAT |
YDR375C | AATTGACAAATTCTCTACTGCTAAACACCCCTTTTGTTCCGAGTTCGCCAGGGATA | GGTAGATTGAGGGCACACTTCATTTTTTTTCAAATCGTCCCTTGGTACGTCTACTG | AAATGAAGTGTGCCCTCAATCTACCGTGCTAAGTGGCGCCTTACTACATAACTTGTATATAATAACACACATTACTTGCCTC | CTAGAAAATATGATTAGCGTTCCGTAAGCTGGAGACCATTTTCAAGATCCCAATACAACAGAT |
YLR035C | CACTCATCTTAGCCACAGTGGAGGGTTTACCTATGAA | CCAAAACTATGAAAAGGCGTCTGTTCCTCAAACCGAG | AACAGACGCCTTTTCATAGTTTTGGTGATCGGGTGCACTATCACCACTCAGCACAATAGATTACTCACACATTACTTGCCTC | TCAGGGGCTGTCTGGCGTGAAGAGGCACCATTCATTTGTCGTGGCGATCCCAATACAACAGAT |
YLR412W | TTTTGCGCTTGTGGTGGTAGCTCAATGGCTTACTCGTT | ACAGCAGAAGACCGGTCAAGGTCATTAACTTCGGTATC | ATGACCTTGACCGGTCTTCTGCTGTCATATCATTCACTACATAGTACTTTATAACTAAACTGTAACACACATTACTTGCCTC | TTAGTACTCTATAGCATGTAACGTCAAGTCAGAGAAGGAATTGATGATCCCAATACAACAGAT |
YML089C | CATCGTGAATTAAGTTCCGGAAATGCGTAGAAAG | AGGGAATAACAACAAATTGGAAATGTGCGGCTAG | ATTACGCTAGCCGCACATTTCCAATTTGTTGTTATTCCCTAAAGTTTTCCACATAATAAACCAAGACAACAGGATTGTTTCACACATTACTTGCCTC | TCACGGATCAATTTTATAAAGCATACAACTAAAGAGAAAAGTGTAGATCCCAATACAACAGAT |
YPL033C | CCAACCTTGTCCTCATCAAGTCAAATGGCGAATATGGCTT | GTCAGATTTTTGGGCTCTATTACTAGTGGACCTGTGCTTC | GAAGCACAGGTCCACTAGTAATAGAGCCCAAAAATCTGACGGGTACTACAAATCGAAGTAACAGTGATAAAATTGCTAAGCACACATTACTTGCCTC | TTAAGTAAAAGAGCTTTTGAAACCAGTTATCAAAAGAATATCACCGATCCCAATACAACAGAT |
Table S10. Table of primer sequences for the 96 ORFs
Figures S10a, S10b, S10c. Gel images showing PCR1 products for the 96 ORFs.
Figures S11a, S11b. Gel images showing PCR2 products for all except 8 of the 96 ORFs. The remaining 8 are shown in the gel in Figure S12.
Figure S12. The remaining eight of the PCR2 results are shown in the first 8 non-marker lanes of this gel, which then also contains the first 24 results for PCR3.
Figures S13a, S13b, S13c. Gel images showing the remaining PCR3 products for the 96 ORFs.
Figure S14. Temperature gradients to investigate PCR1 failures for YLR091W, YML097C, YAL058W and YCL050C.
Figure S15. Temperature gradients to investigate PCR1 failures for YDR071C and YHR207C.
Figure S16. Temperature gradients to investigate PCR1 failures for YJL128C, YML088W, YMR018W and YMR278W.
Figure S17. Temperature gradients to investigate PCR1 failures for YBR255W, YAR002W, YIL148W and YJR073C.
Figure S18. Temperature gradients to investigate PCR1 failures for YPL033C.