Table 1.
List of Primers Used for Preparing Molecular Barcode–Containing Libraries and for Sequencing
| Primer | Chromosome: start-end | Forward primer | Reverse primer |
|---|---|---|---|
| Universal sequence | 5′-GGACACTCTTTCCCTACACGACG-CTCTTCCGATCTCTGNNNNNNNNNN-NNATGGGAAAGAGTGTCC-3′ | 5′-GTGACTGGAGTTCAGACGTGTGCT-CTTCCGATCTGAC-3′ | |
| TP53Exon 2_1 | chr17:7579811-7579985 | 5′-GGGTTGGAAGTGTCTCA-3′ | 5′-GGCCTGCCCTTCCAATG-3′ |
| TP53Exon 3_1 | chr17:7579562-7579747 | 5′-ACTGACTTTCTGCTCTTGT-3′ | 5′-TCATCCATTGCTTGGGACG-3′ |
| TP53Exon 4_1 | chr17:7579491-7579623 | 5′-GGTCCTCTGACTGCTCTTT-3′ | 5′-TTCTGGGAGCTTCATCTGG-3′ |
| TP53Exon 4_2 | chr17:7579369-7579569 | 5′-ATGGATGATTTGATGCTGTCC-3′ | 5′-GCTGCCCTGGTAGGTTT-3′ |
| TP53Exon 4_3 | chr17:7579269-7579411 | 5′-CCTGTCATCTTCTGTCCCTT-3′ | 5′-GGCATTGAAGTCTCATGGAAG-3′ |
| TP53Exon 5_1 | chr17:7578428-7578581 | 5′-AACTCTGTCTCCTTCCTCTTC-3′ | 5′-GCTGTGACTGCTTGTAGATG-3′ |
| TP53Exon 5_2 | chr17:7578337-7578547 | 5′-CTGCCCTCAACAAGATGTTT-3′ | 5′-CAGCCCTGTCGTCTCTC-3′ |
| TP53Exon 6_1 | chr17:7578123-7578321 | 5′-CAGGCCTCTGATTCCTC-3′ | 5′-CACTGACAACCACCCTTA-3′ |
| TP53Exon 7_1 | chr17:7577435-7577645 | 5′-CGCACTGGCCTCATCTT-3′ | 5′-GTCAGAGGCAAGCAGAG-3′ |
| TP53Exon 8_1 | chr17:7576990-7577190 | 5′-ACTGCCTCTTGCTTCTCTT-3′ | 5′-CTCCACCGCTTCTTGTC-3′ |
| TP53Exon 9_1 | chr17:7576822-7576950 | 5′-CACCTTTCCTTGCCTCTTTC-3′ | 5′-CCACTTGATAAGAGGTCCCA-3′ |
| TP53Exon 10_1 | chr17:7573863-7574068 | 5′-ATATACTTACTTCTCCCCCTC-3′ | 5′-TCCTATGGCTTTCCAACCTA-3′ |
| TP53Exon 11_1 | chr17:7572889-7573061 | 5′-GACCCTCTCACTCATGTGAT-3′ | 5′-GTGGGAGGCTGTCAGTG-3′ |
| BRAFExon 11_1 | chr7:140481354-140481532 | 5′-TGTTTGGCTTGACTTGACTT-3′ | 5′-TGTCACCACATTACATACTTACC-3′ |
| BRAFExon 15_1 | chr7:140453058-140453256 | 5′-TCATAATGCTTGCTCTGATAGG-3′ | 5′-ATAGCCTCAATTCTTACCATCC-3′ |
| KRASExon 2_1 | chr12:25398169-25398337 | 5′-ATTATAAGGCCTGCTGAAA-3′ | 5′-GTCCTGCACCAGTAATATG-3′ |
| KRASExon 3_1 | chr12:25380240-25380369 | 5′-AGACTGTGTTTCTCCCTTCT-3′ | 5′-CTCATGTACTGGTCCCTCAT-3′ |
| KRASExon 4_1 | chr12:25378535-25378687 | 5′-AGGACTCTGAAGATGTACCTATG-3′ | 5′-CAGTGTTACTTACCTGTCTTGTC-3′ |
| NRASExon 2_1 | chr1:115258669-115258832 | 5′-TCGCCAATTAACCCTGATTAC-3′ | 5′-ACCTCTATGGTGGGATCATATT-3′ |
| NRASExon 3_1 | chr1:115256459-115256621 | 5′-CCCTTACCCTCCACACC-3′ | 5′-GATGGCAAATACACAGAGGAA-3′ |
| NRASExon 4_1 | chr1:115252120-115252258 | 5′-AAACAAGCCCACGAACTG-3′ | 5′-GGATCACATCTCTACCAGAGT-3′ |
| Custom sequencing primer | 5′-ACACTCTTTCCCTACACGACGC-TCTTCCGATCTCTG-3′ | 5′-GTGACTGGAGTTCAGACGTGTGCTCT-TCCGATCTGAC-3′ |
Note that universal forward and reverse sequences are appended to the 5′ end of each target-specific forward and reverse primer sequence, respectively, before synthesizing the primers.