Sahara One-Step RT-qPCR Master Mix
- 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
Sahara One-Step RT-qPCR Master Mix 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
Fast PCR and Reverse Transcription
The fast reaction mix requires only 5 minutes for reverse transcription, and can subsequently perform 40 cycles of amplification in 20 minutes.
Amplifies Everything Without Inhibition
Sahara One-Step RT-qPCR Master Mix 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%
Safeguard Against Carry-over Contamination
The mix contains a 90% dUTP/dTTP ratio, allowing amplicons to be selectively degraded. If you later encounter carry-over contamination resulting in false positives/amplification of NTC reactions, switching to the Sahara One-Step RT-qPCR Master Mix with UNG will degrade carry-over contamination.
|Optimal RT Activity Temp||60 – 65 ºC|
|RT Step Duration||5 min|
|Hot Start Mechanism||Aptamer|
|Carry-over Prevention||Compatible, dUTP included†|
|GC Content||Up to 70%|
|Multiplexing||Up to 4-plex|
|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|
† Mix is provided with a 9:1 dUTP/dTTP ratio, allowing amplified fragments to be degraded with Sahara One-Step RT-qPCR Master Mix with UNG.
|dNTPs||0.5 mM dATP/dCTP/dGTP|
0.45 mM dUTP
0.05 mM dTTP
|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|
|Storage Conditions||-20 ºC, or 4 ºC for 6 months. Protect from light.|
The mix ships with an optional ROX reference dye, allowing it to be used with high, low, and no-ROX qPCR instruments.
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...
To promote collaboration in science, Sahara One-Step RT-qPCR Master Mix 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