10 seconds and 62 u00b0 c 15 seconds amplification of the wild type hbv dna is inhibited by 12 7 cycles or about 5000 fold while amplification of the mutant the gtg variant is reduced by only 0 39 cycle wi pcr for the hbv bcp region the wi oligo and the amplification primers a and b are 5 u2032 aggagattaggttaaaggtctttgt ph 5 u2032 gaggagttgggggaggagattaggttaa 3 u2032 and 5 u2032 gaagtggtgttcaatttatgcctacagcctccta 3 u2032 respectively capital letters indicate lna nucleotides u201c ph u201d stands for 3 u2032 end phosphorylation the wi pcr is carried out using the hot start dna polymerase 200 u03bcm dntp 0 5 u03bcm each of the amplification primers 2 u03bcm of inhibitory oligonucleotide and the template dna the thermal profile is 95 u00b0 c for 2 10 min to activate the polymerase followed by 18 cycles for downstream qpcr or 45 cycles for dna sequencing of 95 u00b0 c 10 seconds 75 u00b0 c 15 seconds 57 u00b0 c 10 seconds and 65 u00b0 c 5 seconds amplification of the wild type hbv dna is inhibited by 12 68 cycles or about 7000 fold while amplification of the mutant 1762t 1764a is reduced by only 0 24 cycle wi pcr for the precore region of hbv genome the wi oligo and the amplification primers a and b are 5 39.