Optimization of PCR conditions for amplification of GC-Rich EGFR promoter sequence.

ABSTRACT:

Background

Polymerase chain reaction (PCR) is an extremely sensitive method that often demands optimization, especially when difficult templates need to be amplified. The aim of the present study was to optimize the PCR conditions for amplification of the epidermal growth factor receptor (EGFR) promoter sequence featuring an extremely high guanine-cytosine (GC) content in order to detect single nucleotide polymorphisms -216G>T and -191C>A.

Methods

Genomic DNA used for amplification was extracted from formalin-fixed paraffin-embedded lung tumor tissue and PCR products were detected by agarose gel electrophoresis.

Results

Results showed that addition of 5% dimethyl sulfoxide (DMSO), as well as DNA concentration in PCR reaction of at least 2 μg/ml, were necessary for suc

SUBMITTER: Obradovic J 

PROVIDER: S-EPMC6807403 | biostudies-literature | 2013 Nov

REPOSITORIES: biostudies-literature