Comparison of Several Hot-Start Taq DNA Polymerases for Detection of Differentially Expressed Genes by GeneFishing

摘要

GeneFishing (Seegene) is a new proprietary method for detecting differentially expressed genes in two or more related samples. This two-step reverse transcription (RT>PCR method modified from differential display PCR uses arbitrary primer pairs (annealing control primers, ACPs] at the PCR stage with a constant reverse primer (anchor ACP-T) which is also employed to prime the RT reaction. These ACPs have arbitrary 3'-end structures and constant 5'-end structures with an annealing regulatory region in between. With careful adjustment of cycling conditions, it is possible to allow only those cDNAs bearing complementary sequences to the variable 3'ends of the forward primers within a 5'-end region of approximately 2,000 nucleotides to be amplified.