ForeDirect RT-qPCR Kit
Kit Description:
◮ For RNA direct real-time amplification from swab collections without RNA purification processes. This kit completes the qRT-PCR cycles within one hour. The enzyme mix is an optimized blend of Reverse Transcriptase, Hot-Start Taq DNA polymerase, RNase inhibitor. The Reaction Buffer contains all of the required components, including optimized buffer components, Mg2+, dUTP, and dNTPs.
Description
ForeDirect RT-qPCR Kit is designed for RNA direct real-time amplification from swab collections without RNA purification processes. This kit completes the qRT-PCR cycles within one hour. The enzyme mix is an optimized blend of Reverse Transcriptase, Hot-Start Taq DNA polymerase, RNase inhibitor. The Reaction Buffer contains all of the required components, including optimized buffer components, Mg2+, dUTP, and dNTPs.
Specifications
100T
Kit components
| Nucleic acid release agent |
| RNA protectant |
| 2× RT-qPCR Buffer |
| Enzyme Mixture(Taq&M-MLV) |
|
Instructions |
PCR Application
1. Once the PCR reaction tubes have been briefly centrifuged, place them in the sample tank of amplification instrument.
2. Thermal protocol
|
Step |
Temperature |
Time |
Cycles |
|
|
1 |
Reverse transcription |
50℃ |
15 mins |
1 |
|
2 |
Pre-denaturation |
95℃ |
1 min |
1 |
|
3 |
Denaturation |
95℃ |
10 seconds |
42 |
|
4 |
Anneal/Extension |
60℃ |
30 seconds |
|
Note: Fluorescent signal was detected immediately after the extension step of each cycle.
3. After setting, save the file and run the reaction program.
Operating Steps
Before reagent preparation, all the reagents in the kit should be thawed at room temperature and mixed gently.
Reagent preparation
1. Prepare the Nucleic Acid Release Mix
|
Component |
Volume per Sample or Control |
|
Nucleic acid release agent |
5μL |
|
RNA protectant |
0.5 μL |
2. Prepare the Reaction Mix
|
Component |
Volume per Sample or Control |
|
2× RT-qPCR Buffer |
15 μL |
|
Enzyme Mixture(Taq&M-MLV) |
1.5 μL |
|
Primers and Probes |
1μL |
Storage and Shelf life
-The kit should be stored at -20 ± 5℃. The validity period is 12 months.
-Avoid repeated freeze-thaw cycles (<5 cycles).
