Product Description: WISENT ADVANCED 1-Step RT-PCR MIX uses the latest developments in reverse transcriptase technology and buffer chemistry for efficient cDNA synthesis and PCR in a single tube. Proprietary small-molecule hot-start technology offers improved specificity and sensitivity compared to other methods. Our Taq HS DNA Polymerase performs consistently well on a broad range of templates, including both GC-rich and AT-rich ones. We use a thermostable reverse transcriptase working at 45°C to 55°C, along with an advanced RNase inhibitor. Take advantage of our proprietary hot-start technology for unrivalled detection of low copy number templates.
Ordering Information:
CAT.NO. | Quantity | Specification |
801-101-ZR | 50 REACTIONS | RT PCR - 1 STEP |
801-101-AR | 100 REACTIONS | RT PCR - 1 STEP |
801-101-ER | 500 REACTIONS | RT PCR - 1 STEP |
REMARKS:
1.Primers should have a predicted melting temperature of around 60°C using default primer 3 settings.
2.Reverse transscription: an incubating temperature of 45°C . For 10 minutes is recommended except where region of interest contain high secondary structure incubation in which case up to 55°C might be used. For aplicons above 1 kb, the incubation time should be increased to 20 min.
3.Determine the optimal annealing temperature experimentally by performing a temperature gradient.
4.Optimal extension is achieved at 72°C. Optimal extension time depends on amplicon length and template. Normally 15 seconds per kilobase is used for amplification for aplicons between 1 kb and 3 kb.
Product Stability And Storage:
On arrival store at -25°C to -15℃.
Avoid prolonged exposure to light.
If stored correctly the mix will retain full activity for 12 months. It is stable at 2-8°C for one month and can withstand 30 freeze/thaw cycles without any loss of activity.
