Overview
The use of reverse transcriptases to perform first-strand cDNA synthesis has had a huge impact on the study of biological processes. Finally, an RNA could be converted into DNA, allowing manipulation and cloning. Another
important milestone in the evolution of molecular biology was the widespread application of PCR to amplify DNA. It didn’t take long before firststrand cDNA synthesis and PCR were connected and RT-PCR was born. Today, this technique is used almost as often as PCR. Many options are available to researchers, and questions must be answered before deciding which system to choose.
Access RT-PCR System– Maximum Control
AccessQuick™ RT-PCR System– Maximum Convenience
AccessQuick™ and Access RT-PCR Systems
The Access and AccessQuick™ RT-PCR Systems offer the quickest route for amplifying a single, specific message. Both the Access and AccessQuick™ Systems are one-tube, one-step, coupled RT-PCR Systems.
Combine the RNA and gene-specific primers with the kit components, and the entire reaction is ready. The Access System allows the most control over the reaction conditions, and the AccessQuick™ System offers the most convenience. Both use the robust AMV Reverse Transcriptase and the versatile Tfl DNA Polymerase along with a unique buffer that allows the most activity of both enzymes in the same reaction mixture. AMV RT is more heat-stable than M-MLVRT, thus allowing RT steps at temperatures of 42°C or higher. Tfl DNA Polymerase is indistinguishable from Taq DNA Polymerase in reaction characteristics. Like Taq DNA polymerase,
Tfl DNA Polymerase leaves 3′ A overhangs, allowing for direct cloning of amplimers with PCR cloning vectors like the pGEM®-T Easy Vector System.
Comparison of the Access and AccessQuick™ RT-PCR Systems. Human protein phosphatase I (Panel A) and human γ-actin (Panel B) were amplified using the standard methods recommended with each system.
