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Description

Genotyping reaction was prepared in a 10 L reaction containing ~ 5 ng of gDNA, 250 nM K408R Genotyping Forward primer (5'- CCACCATTCTAAATCTACCTAACCTAATA-3' ), 250 nM K408R Genotyping Reverse primer (5'- GGATCGGGCAGTGGTTTTA-3' ), 125 nM wild type Locked Nucleic Acid (LNA) probe ( HEX-TCT + A + A + GT + CT + TCT + GGC-IowaBlack FQ), 125 nM K408R LNA probe ( FAM-TCT + A + G + GT + CT + T + CT-IowaBlack FQ), and 2X PerfeCTa qPCR Tough Mix (QuantaBio)