COSMO RT-PCR Master Mix.

The COSMO RT-PCR kit is designed specifically for simple and effective one-step RT-PCR. The reverse transcription step is directed by M-MuLV reverse transcriptase, converting RNA into complementary DNA (cDNA).

The modified MMLV reverse transcriptase (RTase) is both thermostable and extremely active. The enzyme is blended with RNAase inhibitor preventing degradation of RNA by RNAase contamination, increasing cDNA yield. In addition to increased enzyme kinetics, the RTase is not inhibited by ribosomal and transfer RNAs.

Once the cDNA has been synthesised, a chemically modified ‘Hot Start’ Taq DNA polymerase is activated by heating to 95°C for 2 to 5 minutes, followed by regular PCR. The ‘Hot Start’ DNA Polymerase Enzyme has been isolated from a recombinant E. coli strain containing the DNA polymerase gene from Thermus aquaticus Y-T1 and has been modified to prevent non-specific targets with the addition of antibodies in the buffer.

An optimised reaction buffer provides superior performance for both reverse transcriptase and Taq DNA polymerase allowing you to easily convert your RNA into cDNA for amplification followed by PCR in a single step. This product provides you with an all-in-one protocol, increase efficiency and accuracy. All that is required is your DNA template and gene-specific primers.

For further information, please see below or contact us at info@willowfort.co.uk

Features

  • Efficient – Improves yield by performing reverse transcription and PCR in a single tube, thus reducing cDNA loss.
  • Accurate – Minimises the production of non-specific targets by including monoclonal antibodies which control ‘Hot Start’ Taq DNA polymerase enzyme activity.
  • Robust– High fidelity amplification with little optimisation required and amplifies products up to 9 kilobases.

For research use only. These products should not be used for diagnostics.

Use the tables below as a quick guide on how to use the COSMO RT-PCR Master Mix product:

Table 1: Reagents required per sample

Components

Amount

COSMO ‘Hot Start’ PCR RED Master Mix

25μl

Forward and Reverse Primers

1-2.5μl (0.1-1μM each)

Nuclease-free water

To 50μl

DNA template (10pg—1μg)

Volume varies

 

Table 2:  Temperature and Times for the PCR protocol

Stage

Temperature

Time

Stage 1

95°C

*2 min

Stage 2.1

95°C

10 sec

Stage 2.2

Primer Tm <5°C

20 sec

Stage 2.3

72°C

#30-60s

Repeat stage 2 for 25-35 cycles

Stage 3

72°C

1 min

*Increase to 5min if GC content is higher than ≤50%

# The extension time depends upon the amplicon length

 

For more information about the COSMO RT-PCR Master Mix protocol, please click here for the guide.

To place an order, please contact us through the “Contact Us” link, quoting the Catalog No.

Product

Package Size Catalog No.
COSMO RT-PCR Master Mix 25 reactions

WF10204001

100 reactions

WF10204002

Storage Conditions

Upon delivery of the COSMO RT-PCR Master Mix, store all reagents in -20°C.

Troubleshooting Guide

Please click here for the PCR Troubleshooting guide.

If you require any additional information about this product please email us at info@willowfort.co.uk