Page 58 - TD-3-3

Tumor Discovery Somatic mutations in POLE and POLD1 in colorectal cancer

Appendix

Table A1. The outer primer sequences
Gene/Exon Forward Reverse Amplicon size
POLE/Exon 9 5’ATGGGGAGTTTAGAGCTTGG’3 5’CCATCCCAGGAGCTTACTTC’3 192 bp
POLE/Exon 10 5’GCAGCCTCTGACTTGTGC’3 5’CCGTTCTTCCCACAATACC’3 200 bp
POLE/Exon 11 5’GCAGACCTCTGACTGCTGTG’3 5’CCCTCCCTCTCAAATGCTG’3 167 bp
POLE/Exon 12 5’GGCATTAGAGCCTGACCTG’3 5’CACGGACAGCAGTGAGGAG’3 214 bp
POLE/Exon 13 5’CGGGCTGCATGTTAGAATC’3 5’AGCCGGGATGTGGCTTAC’3 205 bp
POLE/Exon 14 5’TTTGATGGCCCTGCTCTC’3 5’TCCATTCAGCTCCAGTGC’3 236 bp
POLD1/Exon 8/A 5’ CTGGGAAATACGCCCTGAG’3 5’ CGATATCGAAGCTGAGCAC’3 220 bp
POLD1/Exon 8/B 5’ ACGTGCTGTGGTCTGACG’3 5’ CCTTCATGCTTGCCATTCC’3 201 bp
POLD1/Exon 9 5’ AGCCTCCCTGCTGTGTTG’3 5’ GGAGCTGATGGCTCAGGAC’3 304 bp
POLD1/Exon 10 5’ CACCAGGGTGACCCAATG’3 5’ GAGGCACACAGAGCAAGATG’3 255 bp
POLD1/Exon 11 5’ TGTGGTTGGTCTCAATCTCC’3 5’ CCTTCCAAGGACAGGGACAC’3 263 bp
POLD1/Exon 12 5’ GGGTGTGTCCCTGTCCTTG’3 5’ CAGCAGGTCAAAGAGTGAGG’3 293 bp
POLD1/Exon 13/A 5’ AGGCTACCTCACCCTGACC’3 5’ GCCACGACTGAGCAGGTAG’3 247 bp
POLD1/Exon 13/A 5’ATGCCTACCTGCCACTGC’3 5’GGATGCCATTGCCTGGAG’3 215 bp
Note: Two sets of primers were used to amplify exon 8 and exon 13 of the POLD1 gene.

Table A2. The nested primer sequences

Gene/Exon Forward Reverse Amplicon size size
POLE/Exon 9 5’TGCTTATTTTGTCCCCACAG’3 5’TACTTCCCAGAAGCCACCTG’3 153 bp
POLE/Exon 10 5’CCTCTGACTTGTGCTGATTGC’3 5’CCGGGTAGTTTCCCAAGTG’3 179 bp
POLE/Exon 11 5’CTGCTGTGACTTGGGTTCAG’3 5’CAAATGCTGCCCAGTTACTC’3 133 bp
POLE/Exon 12 ’TGTCTGTCCTCTTTCCAACC’3 5’AGCCATGCTGCTCTGTGG’3 153 bp
POLE/Exon 13 5’TGGCTTCTGTTCTCATTCTCC’3 5’CGGGATGTGGCTTACGTG’3 179 bp
POLE/Exon 14 5’CTCTGGCGTTCTCTCCTCAG’3 5’GCCGACAGGACAGATAATGC’3 158 bp
POLD1/Exon 8/A 5’TGAGGCTGAAGGAGAAGGTG’3 5’CTTCCGGTGGGTGACTGAC’3 157 bp
POLD1/Exon 8/B 5’GCTGTGGTCTGACGTGGTC’3 5’GCTTGCCATTCCCTCCAC’3 190 bp
POLD1/Exon 9 5’GTGAGGGGCAGGAGTCAG’3 5’GCGTGGAGCGAGAGCTAC’3 228 bp
POLD1/Exon 10 5’AGGGTGACCCAATGTGCTC’3 5’GAGAGAAGCCTCCCACCTG’3 165 bp
POLD1/Exon 11 5’CACACCCTGCCTCTCCTC’3 5’CCAAGGACAGGGACACACC’3 200 bp
POLD1/Exon 12 5’GTCCTTGGAAGGCCACTG’3 5’ GTCAGAGGTTGGGGTGAGAG’3 207 bp
POLD1/Exon 13/A 5’CCTTCTCCTGCTCCACCTC’3 5’CAGTGACCCTCGCCATCTC’3 189 bp
POLD1/Exon 13/B 5’TCATGGTGCTGGTGAACG’3 5’GACAAGTCTCGGCTACTGACC’3 130 bp
Note: Two sets of primers were used to amplify exon 8 and exon 13 of the POLD1 gene.

Volume 3 Issue 3 (2024) 10 doi: 10.36922/td.3659