5 × Hifair™ RT-qPCR Buffer _ 16630ES-A

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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 16630_MSDS_HB251209_EN.PDF Manuals 16630_Manual_Ver.EN20251209.pdf Show more