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Table 2 The HRM primers were designed with Methyl Primer software to screen for methylation of a total of 61 CpG sites in the HPV genome, based on the results of the integration experiments, in order to avoid breakpoints in each sample

From: Identification of reliable biomarkers of human papillomavirus 16 methylation in cervical lesions based on integration status using high-resolution melting analysis

Primer Sequence (5′–3′) CpG sites Size (bp)
E2BSs F-AATTTATGTATAAAATTAAG nt31, nt37, nt43, 106
R-CCTAAAACATTACAATTCTCTT nt52, nt58
E2 F-TTGAAATTATTAGGTAGTATTTGGTT nt3412, nt3415, nt3417, nt3433, nt3461, nt3462  
R-TAATYGTCTATATTTCTTYGATG 90
L1-1 F-GTTTTTTGTAGATTTAGATTAGTTTTTTT nt7034, nt7091, nt7136, nt7145 187
R-AAACAACACTAATTCAACATACATACAA
L1-2 F-TGTTAGAATTATATGGCGATAGTT nt6367, nt6389, nt6457 124
R-CCTTTAATATATAAATCGTCTAATACATT
L1-3 F-TAATTTAGATATATAGCGGTTGGT nt5927, nt5963 81
R-CTAATACCCACACCTAATAACTAA
L1-4 F-GAGTATAGGGTTATAATAATGGT nt6581, nt6650 109
R-AAATAAATATAACAACACATAATAACAT
L1-5 F-TTAAGGAGTATTTACGATATGGG nt6731, nt6796 199
R-ACAATTACCTAAATTACAAACCTATA
L1-6 F-TTTGATATATTTATTAATATAATTGATTA nt5600, nt5606, nt5609, nt5615 142
R-CYTTTACYTCYTTTTCYTAACATATA
L1-7 F-TGTAAGTACGGATGAATATGTT nt5709, nt5726 81
R-AAATATCCAACTACAAATAATCTAAA
L2-1 F-AATAAATTGTTATTATTTAATAATGCGAT nt4240, nt4249, nt4261, nt4270, nt4277 120
R-ATAATATCAAATAAACATATACCTACCTA
L2-2 F-TTTGATATTATATTTAAGGTTGAAGGT nt4427, nt4437, nt4441 125
R-CCAATACGTCCGCCTATAC
L2-3 F-ATAGTAATTAGTAGTATATTTATATTAGG nt4894, nt4906, nt4924 90
R-TACAACTTTAACCTATTATATTATACG
L2-4 F-TTAGGCGTATTGGTATTAGGT nt5128, nt5173, nt5179 101
R-ATCATAATAATAATATACCTTAACACCT
L2-5 F-TTTTATAATTTYGGTAGGATTTGTA nt5378 150
R-CTTAATCAATTATATTAATAAATATATCAA
E6 F-TTGTTAATTAGGTGTATTAATTGTTAA nt494, nt502, nt506, nt539 179
R-TATATCTCCATACATAATTACAACTAA
E7 F-GATAAGTAGAATCGGATAGAGTT nt701, nt752, nt757, nt765, nt780, nt789 137
R-CCTAATATACCCATTAACAAATCTT
Clone-LCR F-CCTACTAATTGTGTTGTGGT   689
R-CTCCTGTGGGTCCTGAA
Clone-E6 F-GGAACAACATTAGAACAGCA   231
R-GCAATGTAGGTGTATCTCCA
Clone-E7 F-GCAACCAGAGACAACTGA   231
R-GGGCACACAATTCCTAGT
Clone-E2 F-AGCAACGAAGTATCCTCTC   187
R-TGTCCACTGAGTCTCTGT
Clone-L2 F-CCTCTGCGTTTAGGTGTT   1504
R-GAAGGAGCTTGGTCAGTTAT
Clone-L1 F-TTCCAGGGTCTCCACAATA   1693
R-GCACATACAAGCACATACAA
  1. Primers for high-resolution melting PCR for determination of human papillomavirus 16 methylation and construction of standard curves