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

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):

[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

16630_MSDS_HB251209_EN.PDF

Manuals

16630_Manual_Ver.EN20251209.pdf