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. 2017 Sep 27;15(3):208–212. doi: 10.15171/ijb.1307

Table 1. PCR primers and conditions. F1 and R1 (outer primers), F2 and R2 (inner primers) were designed for T-ARMSPCR. To enhance the specificity of inner primers (F2 and R2), at the 3rd nucleotide from the 3´-terminus changes to destabilizing mismatch (the bold underlined bases indicate the extra mismatched bases introduced). F3 and R3 primers were used for PCR- RFLP reaction.

Primer sequence Tm (ºC) Amplicon size (bp)
F1: 5'-AGCCTCGGTGAGATAAAGGATG-3'
R1: 5'-CCTGGACCTGCTCTGATTGTC-3'
F2: 5'-GCTGCTGCAGGCCCCAGATAAG-3'
R2:5'-GCAGAAGGAAGAGTTCTGGGAGA-3'
F3: 5'-TCACGGAGACCCAGCCAATGAG-3'
R3: 5'-TCCATCCCACCCAGTCAATCCC-3'
66
66
72
70
70
70
G allele (475)
T allele (271)
Control band (701)

G allele undigested (293)
T allele digest with MboI (197+95)