Extended Donor Typing by Pooled Capillary Electrophoresis: Impact in a Routine Setting.

ABSTRACT:

Background

PCR with sequence-specific priming using allele-specific fluorescent primers and analysis on a capillary sequencer is a standard technique for DNA typing. We aimed to adapt this method for donor typing in a medium-throughput setting.

Methods

Using the Extract-N-Amp PCR system, we devised a set of eight multiplex allele-specific PCR with fluorescent primers for Fy^a/Fy^b, Jk^a/Jk^b, M/N, and S/s. The alleles of a gene were discriminated by the fluorescent color; donor and polymorphism investigated were encoded by product length. Time, cost, and routine performance were collated. Discrepant samples were investigated by sequencing. The association of new alleles with the phenotype was evaluated by a step-wise statistical analys

SUBMITTER: Wagner FF 

PROVIDER: S-EPMC6158588 | biostudies-literature | 2018 Jul

REPOSITORIES: biostudies-literature