 Transcribing In Reverse
Sep 17 '07
As a tool used in the diagnosis of genetic diseases and in evaluating gene expression, RT-PCR (reverse transcriptase-polymerase chain reaction) has become a very common and useful biological technique. Similar to PCR kits and polymerases, there is an abundance of companies and products available. One of the main differences between kits is whether the kit offers a one-step versus a two-step RT-PCR reaction. (In a two-step reaction, the RT and PCR steps are set up sequentially, while in a one-step reaction, all components are combined initially so that there is no user intervention between the RT and PCR reactions.). If the RT-PCR reaction has already been optimized, a generic quick one-step kit is definitely the easiest, preferred method. You will pay a little less, but spend more time setting up a two-step reaction. A two-step reaction might be required for more challenging RNA samples and PCR conditions, especially if RNA abundance is low. Two-step RT-PCR is also useful if there are multiple PCR reactions to be performed with the cDNA produced in the first step. When troubleshooting is required or the reaction has special issues, there are various products out there that promise to help out. There are kits that can be used for Long Range RT-PCR (usually requiring two-steps), different enhanced versions of the RT enzyme, several Taq polymerases promising jump start, hot start, increased specificity, robustness, and low error rates. There are numerous RT and Taq polymerase variants available, with each company claiming an advantage or benefit for its products over the others. As always, start simple before getting into products with more complicated features which may not be necessary, and keep in mind the reputation of the company.
Elizabeth Hawkes
Research Associate
University of California, San Francisco
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| Company |
Invitrogen |
Promega |
| Item |
SuperScript™ III One-Step RT-PCR System with Platinum® Taq |
AccessQuick™ RT-PCR System |
| Price |
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| Description |
The SuperScript™ III One-Step RT-PCR System with Platinum® Taq DNA Polymerase offers the highest sensitivity available for end point detection. This system combines the high thermostability of SuperScript™ III RT with the specificity of Platinum® Taq DNA Polymerase to increase priming specificity, generate higher product yields, and identify a larger range of targets...
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The AccessQuick™ RT-PCR System(a) is an easy and convenient master mix system for setting up one-tube RT-PCR reactions. It is designed to increase the convenience of performing RT-PCR by combining the following components in a single tube: Tfl DNA Polymerase, dNTPs, magnesium chloride and reaction buffer...
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| Catalog Number |
12574018 |
A1701 |
| Quantity |
25 Rxn |
20 reactions |
| Kit Contents |
SupersriptTMIII RT/platinumTaq mix -50 micro liters
2X reaction mix ( a buffer containing 0.4 mMof each dNTP,3.2 mMMgSO4-1ml
5mM MgSO4-500 micro liters |
2 x 250µl AccessQuick™ Master Mix (2X), 1 x 100u AMV Reverse Transcriptase, 1 x 1.25ml Nuclease-Free Water
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| RT Type |
One-Step |
AMV Reverse Transcriptase |
| Comments |
This kit has been optimizedfor end point of RT-PCRThe convenient 2X format allowto addtemplate & primer at any desired concentrationAutomatic "hot start" in PCR for incres sensitivity,specificity&yield |
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