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