Table 3. L2. Primers sequences and annealing temperatures.
Target Region | HPV type | PCR primers and assay conditions | Pyrosequencing primers | |||
---|---|---|---|---|---|---|
PCR primer a | Ampl. bp | PCR ann. T |
Sequencing a primer | HPV strand | ||
L2 b | 16 | Forward GTTAGGTGGATATGTATTTGTT Reverse bio- ATACCATTATTTTTAATACATACACATAC |
276 | 58 | ATGTTTTATAAAGTTGGG | Lower |
18 | Forward AGGTTGGTTTATATAGTGTATTGT Reverse bio- ATACCTATACCAAATCCACC |
263 | 56 | TTGTATTTTTATAATAAAATTATGG | Upper | |
31 | Forward TGGATAAGTATTTGTTGTTTTATATGTT Reverse bio- CCATTATTTATAATTCATACACATACAT |
235 | 50 | TTTGATATAATTGTGTAGTAG | Lower | |
33 | Forward GGTTTTGTATGTTTGGTATAGTT Reverse bio- ACTAATAAATACCTTTATATTTTAACAAT |
106 | 50 | GTATAGTTGTGTTGTAGATG | Lower | |
35 | Forward TGGATAAGTTTTTGTAGTTTTGTAA Reverse bio- AATAAATTAATCACATAATATAACCATAC |
112 | 48 | TAGTTGTGTTGTAGATG | Lower | |
39 | Forward bio- ATAGGTTTTATATAGGTTAGTTGTAGA Reverse ACCACACCATTAAAAATATTTTATA |
226 | 56 | AATAAACATAATTTCCCACC | Lower | |
45 | Forward bio- TATGTTTTATATAAGTTAGTTGTAGA Reverse ACATCCCCATTAACAACATTTACTA |
269 | 56 | AACCATAATATCCCACC | Lower | |
51 | Forward GTTTTTGGTTTGTTGTTGTAATATTTTAA Reverse bio- AACCCACTCCACTATAATATTTTATC |
87 | 56 | TTGTGATTAAATATGGTGG | Upper | |
52 | Forward GTTAGAGGTTTTGTATGTTTGATATAG Reverse bio- ATTTTCCAATATTTTATATTCACTATCAT |
239 | 50 | ATATAGTTGTGTTGTAGAA | Lower | |
56 | Forward TTGGATATGTATTAGATAATTTATATGTT Reverse bio- ACCACATCCTTTTTTAATACATTTATA |
209 | 48 | ATATAGTTGTGTTGTAGA | Lower | |
58 | Forward GGTTTTGTATGTTTGGTAAAGTT Reverse bio- CATACACAATACTTAACCCAAC |
221 | 58 | GGTAAAGTTGTGTAGTAGA | Lower | |
59 | Forward GGGTATGTATTTGTTTGTTTGTAG Reverse bio-AATACTATATACCCATACAATACTATCC |
221 | 56 | AAGTTTGTTGTTGAGG | Lower |
a Bisulfite modified sequences
b L2. HPV L2 gene. Reference HPV16 CpG: 4261