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Table 1 Validation of MS-MLPA results for ATM, MLH1 and FHIT

From: Promoter methylation of tumor suppressor genes in pre-neoplastic lesions; potential marker of disease recurrence

Gene Method Primer sequence/polyclonal antibody No. samples examined Overall concordance (%)
ATM Pyrosequencing CpG analysis Fw: 5′-AGAAGTGGGAGTTGGGTAGTT-3′ 77/78 73%
Rv: 5′-biotinCTCCCCCCCCCTACCACTACACTC-3′
Seq: 5′-AGGAGGAGAGAGGAGT-3′
MLH1 Pyrosequencing CpG analysis Fw: 5′-biotinGGGAGGTAAGTTTAAGTGGAATAT-3′ 72/78 79%
Rv: 5′-CCAATCCCCACCCTAAAACCCTC-3′
Seq: 5′-CTAAACTCCCAAATAATAACCT-3′
FHIT Immunohistochemistry Rabbit polyclonal anti-FHIT; clone PA1-37690; Thermo Scientific Pierce; working dilution: 1/200 57/78 84%
  1. Abbreviations: Fw Forward Primer, Rv Reverse primer, Seq sequence analyzed.