qRT-PCR is an invaluable tool for studying gene expression. The method relies on the central dogma of Molecular Biology, where DNA is transcribed into messanger RNA (mRNA) which is then translated into protein. From a large population of total cellular RNA, the mRNA of interest is selectively reverse transcribed to generate a complementary DNA (cDNA). In turn, using a pair of specific primers, the cDNA is amplified. There are two alternative approaches to quantify the gene expression in real time: First, using fluorescent SYBR Green dye which intercalates between the bases of double stranded DNA and the intensity of the fluorescent signal is directly proportional to the extent of DNA amplification. Alternatively, TaqMan chemistry relies on a gene-specific oligonucleotide probe with a fluorophore and a quencher molecule at either ends in addition to a pair of specific primers. As DNA amplification occurs, the fluorescence signal increases in a proportional manner.
At o2h, we offer both TaqMan® as well as SYBR Green® based comprehensive gene expression analysis services.
To know more about our biology services offering or to request our brochure, please reach out to us at discovery@o2h.com.
