Features/Description:
Description:
RNAScribe RT Reverse Transcriptase (RT) is a genetically engineered MMLV (Moloney Murine Leukemia Virus) based Reverse Transcriptase. This is an RNA‐directed DNA polymerase that can synthesize a complementary DNA strand from ssRNA or ssDNA and is active over a broad range of reaction temperatures from 37°C-60°C. RNAScribe RT is a robust enzyme for RNA detection and has enhanced stability at room temperature with no activity loss for up to 1 month. This RT contains a functional RNase H domain which can increase the sensitivity of RT‐qPCR (quantitative reverse transcription PCR).
Applications:
Limit of use:
max. Target length: 8 kb
Kit Components for RT-qPCR:
1. BioStar Enzyme-mix (25x)
50 mM Tris-НCl (рН 8.0 at 25 °С), 100 mM NaCl, 1 mM EDTA, 5 mM DTT, 50 % (v/v) glycerol and 0.1 % (v/v) NP-40, RNAscribe RT reverse transcriptase, highly-processive HS-Taq DNA polymerase and inhibitor of RNases.
2.) Reaction Buffer (2x):
100 mM Tris-НCl (рН 8.3 at 25 °С), 150 mM KCl, 0.6 mM each dNTP, 8 mM MgCl2, 10mM TCEP, stabilizer
Storage:
@ -20°C, short term at room temperature up to 6 days.
@ +4°C for about 10 days.
About 30 times Multiple thawing/freezing without any loss of activity.
Transport:
the product will be shipped with "blue ice"; up to 6 days @ room temperature.
Protocols / Manuals / 25 µl total mix
The datasheet includes detailed protocols for your assay.
- Defrost and mix all components and spin gently
- Add the volumes from the table to thin- wall PCR tubes (preferred on ice)
Component | Volume | final. conc |
2x Re-action mix | 12,5 µl | |
25x Bio-Star Mastermix | 1,0 µl * | |
Forward primer | variable |
0,1 - 500 nM |
Reverse Primer | variable |
0,1 - 500 nM |
Probe | variable |
0,1 - 500 nM |
RNA template | variable | 1 pg - 1 µg |
Sterile water | up to 25 µl |
* range from 1-3 µl
- Centrifuge your reaction chambers
- Start your PCR-Cycler routine:
Step | Temp. °C | Time | cycles |
Reverse Transcription |
45-55 | 10-15 min | 1 |
1. Denaturation | 94 | 3-5 min | 1 |
Denaturation | 93-94 | 5-15 sec | 25-45 |
Annealing | 50-68 (Tm-5) | 5-20 sec | 25-45 |
Elongation | 72 | 0,5 min/kbp | 25-45 |
RT-qPCR Mastermix, RT-PCR Mastermix with unique RNAScribe RT reverse transcriptase and HotStart Taq DNA Polymerase
Final price excl. shipping costs3