Descrição
5 × Hifair™ RT-qPCR Buffer is the buffer component of the Hifair™ Multiplex One-Step RT-qPCR Probe Kit (UDG Plus) (Cat# 16630). When combined with the corresponding enzyme mix, it enables multiplex quantitative PCR directly from RNA templates.
In this workflow, reverse transcription and qPCR are performed sequentially in a single tube, streamlining the procedure and minimizing the risk of contamination.
Components
|
Components No. |
Name |
16630ES-A60 (100 T) |
16630ES-A80 (1,000 T) |
16630ES-A92 (10,000 T) |
|
16630ES-A |
5×Hifair™ RT-qPCR Buffer |
500 μL |
5 mL |
50 mL |
Storage
This product should be stored at -25~-15℃ for 1 year.
Notes
1. This product is for research use only.
2. Please operate with lab coats and disposable gloves，for your safety.
Instructions
1. Preparation of reaction system((25 µL Example):
|
Components NO. |
Components Name |
Volume(μL) |
Final Concentration |
|
16630ES-A |
5×Hifair™ RT-qPCR Buffer |
5 |
1× |
|
16630-B |
Hifair™ Enzyme Mix |
1 |
- |
|
16630-C |
GC-Enhancer |
0-5 |
- |
|
/ |
Primer Mix (10 μM) |
0.4 each |
0.16 μM |
|
/ |
Probe Mix (10 μM) |
0.2 each |
0.08 μM |
|
/ |
Tempelate RNA |
1-5 |
- |
|
/ |
RNase Free H2O |
to 25 |
- |
[Note]: Thoroughly mix all reagents before use. Avoid vigorous pipetting or shaking to prevent excessive bubble formation.
1. Primer Concentration:
The Primer Mix contains multiple primer pairs. The final concentration of each primer can typically be adjusted between 0.1–1.0 μM, depending on assay requirements.
2. Probe Concentration:
The Probe Mix includes multiple probes labeled with distinct fluorophores. The final concentration of each probe can be optimized within the range of 50–300 nM.
3. Template Dilution:
Due to the high sensitivity of qPCR, template dilution is strongly recommended to achieve optimal Ct values in the range of 20–35.
4. Reaction Volume:
A reaction volume of 25 μL or 50 μL is recommended to ensure robust amplification and reproducibility of target genes.
5. Preparation of reaction system:
1)Prepare the master mix in a biosafety cabinet or clean workspace.
2)Use nuclease-free pipette tips and PCR tubes.
3)Filtered (aerosol-resistant) tips are highly recommended.
4)Take precautions to avoid cross-contamination and aerosol carryove
Documents:
Safety Data Sheet
Manuals
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