Hifair™ AdvanceFast One-step RT-gDNA Digestion SuperMix for qPCR _11151ES

Hifair™ AdvanceFast One-step RT-gDNA Digestion SuperMix for qPCR represents an enhanced iteration of the Hifair™ Ⅲ 1st Strand cDNA Synthesis SuperMix for qPCR (gDNA Digester Plus). It is designed to fulfill the demands of conducting both reverse transcription and genome removal reactions within a single tube. Moreover, it streamlines the reaction process, significantly reduces reaction time, and augments the yield of the reverse transcription product. The 4×Hifair® AdvanceFast One-Step RT SuperMix encompasses all essential reagents for the reverse transcription reaction. During the reaction, you simply need to supplement it with the gDNA Remover Mix, template RNA, and water to efficiently synthesize the initial strand of cDNA while concurrently eliminating genomic DNA contamination. This product is well-suited for the reverse transcription of intricate RNA templates and proves highly effective for reverse transcription involving scant template amounts and low-copy genes.

The reverse transcription product generated by this particular product is fully compatible with both the probe-based and dye-based qPCR methods. For probe-based qPCR, we recommend using Hieff Unicon® Universal TaqMan multiplex qPCR master mix (Cat#11211), while for dye-based qPCR, Hieff UNICON® Universal Blue qPCR SYBR Green Master Mix (Cat#11184) is the recommended choice for high-performance gene expression analysis.

However, it's important to note that the reverse transcription product produced by this product is not suitable for subsequent applications such as the amplification of long fragments. If you require efficient reverse transcription for these purposes, we recommend utilizing the Hifair® AdvanceFast 1st Strand cDNA Synthesis Kit (Cat#11149, Cat#11150).

Features

• 5 mins of RT

• Integrated RT and gDNA digestion in one tube

Applications

• RT-qPCR
  
• Antisense RNA synthesis

Specifications

Components

No.	Name	11151ES10 (10T)	11151ES60 (100T)
11151-A	4×Hifair® AdvanceFast One-Step RT SuperMix	50 μL	500 μL
11151-B	gDNA Remover Mix	10 μL	100 μL
11151-C	RNase-free Water	1 mL	2×1 mL

Shipping and Storage

The product is shipped with dry ice and can be stored at -15℃ ~ -25℃ for 12 months.

Instructions for use

Thaw the 11151-A/B/C components on ice, gently agitate and thoroughly mix each reagent component. Then, assemble the following components within an RNase-free tube:

Component	Vol（μL）
RNase-free Water	To 20 μL
4×Hifair® AdvanceFast One-Step RT SuperMix	5 μL
gDNA Remover Mix	1 μL
Total RNA	10 pg -5 μg*
or mRNA	10 pg-500 ng*

Table 1: Reaction preparation

*We suggest not exceeding 2 μg as the total RNA input amount. In case of low expression abundance of the target gene, the total RNA input can be increased up to a maximum of 5 μg.

Temperature	Time
37℃	5 min*
85℃	30 s

 

Table 2: Reaction Protocols

*This process encompasses both reverse transcription and gDNA digestion.

The reverse transcription product is suitable for immediate use in qPCR reactions or can be stored at -20°C for a short duration. For extended storage, we recommend aliquoting and storing it at -80°C to prevent multiple freeze-thaw cycles.

Precautions:

1. All procedures should be conducted on ice, with utmost care to prevent RNase contamination.

2. Ensure your safety and well-being by wearing a laboratory coat and disposable gloves.

3. This product is intended solely for scientific research purposes and should not be used otherwise.