Molecular genetic diagnostic techniques in choroideremia.

ABSTRACT:

Purpose

To optimize and streamline molecular genetics techniques in diagnosing choroideremia (CHM).

Methods

PCR primers were designed for exons 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, and 15 of the CHM gene. Each PCR protocol was optimized so that all exons could be amplified with the same component ratio and PCR conditions. Sense and antisense primers were tested for their ability to be used as sequencing primers. Fibroblast cells were cultured, and an immunoblot analysis was performed to detect the presence or absence of Rab escort protein 1 (REP-1) in a suspected CHM patient sample when no mutation was detected with sequencing. Multiplex ligation-dependent probe amplification (MLPA) of the CHM gene was performed and used to detect deletions and duplications in affected males and

SUBMITTER: Furgoch MJ 

PROVIDER: S-EPMC4000712 | biostudies-literature | 2014

REPOSITORIES: biostudies-literature