Sahara One-Step RT-qPCR Master Mix with UNG
Highlights
- Eliminate false positives resulting from PCR carry-over contamination
- Multiplex up to four RT-qPCR reactions with no loss of efficiency
- Thermally stable mix withstands 40 ºC for seven days without loss of performance
- Fast 5 minute reverse transcription step
- Overcomes inhibition in crude extracts and environmental samples
- License-free for commercial or diagnostic use
Description
Sahara One-Step RT-qPCR Master Mix with UNG is a high performance mix for multiplex, probe-based quantitativie reverse transcription PCR (RT-qPCR). It amplifies RNA targets at high efficiency while resisting PCR inhibitors. The one-step mix allows cDNA synthesis and subsequent PCR amplification to be performed in a single reaction, maximizing operational efficiency and sensitivity.
Extreme Thermal Stability
Stable at 40 ºC for seven days, or 25 ºC for weeks, Sahara One-Step
Run Ultra-Fast Protocols
The fast reaction mix can perform both UNG digestion and reverse transcription in as little as 6 minutes, and can subsequently perform 40 cycles of amplification in 20 minutes.
Amplifies Everything Without Inhibition
Sahara One-Step RT-qPCR Master Mix with UNG has been validated against mRNA, rRNA, viral RNA, and genomic DNA targets from a wide range of organisms. Numerous enhancers bind & inactivate PCR inhibitors, allowing amplification from crude extracts. The mix amplifies GC-rich targets up to 70%
Prevent Carry-over Contamination
Eliminate false positives caused by PCR carry-over contamination. The mix contains a 90% dUTP/dTTP ratio, allowing amplicons to be selectively degraded. If a RT-qPCR reaction is contaminated with a prior amplicon, it will be degraded by the mix's UNG enzyme during the initial 37 ºC incubation step, preventing false amplification.
Specifications
Concentration | 2X |
UNG Activity Temp | 37 ºC |
UNG Step Duration | 1 – 5 min |
Optimal RT Activity Temp | 60 – 65 ºC |
RT Step Duration | 5 min |
Hot Start Mechanism | Aptamer |
Carry-over Prevention | Yes, 9:1 dUTP/dTTP ratio and UNG included |
GC Content | Up to 70% |
Multiplexing | Up to 4-plex |
Reaction Speed | Fast |
RNase Inhibitor | Yes |
Visible Loading Dye | Yes |
Exonuclease Activity | 5' → 3' |
Supported Probes | TaqMan/Hydrolysis, Molecular Beacon, Scorpions |
Supported Templates | Total RNA, mRNA, rRNA, viral RNA, DNA |
Recommended Reaction Volume | 10 – 50 µL |
Reaction Conditions
dNTPs | 0.5 mM dATP/dCTP/dGTP 0.45 mM dUTP 0.05 mM dTTP |
K+ | 20 mM |
Mg2+ | 4 mM |
Mn2+ | 0.75 mM, adjustable 0.5 – 1.5 mM |
Shipping and Storage
Temperature Stability | 7 days @ 40 ºC, weeks @ 25 ºC |
Freeze/Thaw Cycles | Up to 20 |
Shipping Conditions | Ambient |
Storage Conditions | -20 ºC long-term, or 4 ºC for 6 months |
Compatible Instruments
The mix ships with an optional ROX reference dye, allowing it to be used with high, low, and no-ROX qPCR instruments.
FAQs
What is a PCR Master Mix?
A PCR master mix is a premixed solution that contains most of the components necessary to run a PCR assay. The mix contains Taq DNA polymerase, dNTPs, MgCl2, as well as enhancers and stabilizers in a buffer that is optimized for DNA amplification by PCR. Read more...
License
To promote collaboration in science, Sahara One-Step RT-qPCR Master Mix with UNG is provided under an open license:
- It may be used freely for any application, including commercial and diagnostic use
- No license agreements or royalty payments required for OEM use
- May be incorporated into your own test kits without restriction
Resources
Certificates of Analysis
View Sample Certificate |