RT-PCR on Embryos Using Degenerate Oligonucleotide Primers

Abstract

The polymerase chain reaction (PCR) is an incredibly versatile technique that has made a profound impact on many areas of biology. PCR is based on the use of oligonucleotides, which flank the region of DNA of interest and which are complementary to sequences on either DNA strand, to prime the replication of each strand by Taq DNA polymerase. This enzyme is thermostable, and consequently, one can go through cycles of template denaturation and renaturation after each round of replication and exponentionally amplify the sequence of interest. This procedure is very sensitive, and as such, it allows one to amplify, isolate, and utilize specific DNA sequences from vanishingly small quantities of starting material using either genomic or cDNA as a substrate.