Hifair™ Ⅲ 1st Strand cDNA Synthesis SuperMix for qPCR (gDNA Digester Plus)



Hifair™ Ⅲ 1st Strand cDNA Synthesis SuperMix for qPCR (gDNA Digester Plus) is a readjust premix developed based on Hifair™ⅢReverse Transcriptase. Compared with Hifair™ ⅡReverse Transcriptase, Hifair™ Ⅲ Reverse Transcriptase has significantly higher thermal stability and can withstand reaction temperatures up to 60℃, and is suitable for Reverse transcription of RNA templates with complex secondary structures. At the same time, the enzyme enhances the affinity with templates, which is ideal for the reverse transcription of a small number of templates and low-copy genes.

The premix contains 5×gDNA Digester Mix and 4×Hifair™ Ⅲ SuperMix Plus. 5×gDNA Digester Mix can remove residual genomic DNA contamination in RNA templates to ensure more reliable follow-up results. 4×Hifair™ Ⅲ SuperMix Plus contains all components required for Reverse Transcriptase (Buffer, dNTP, Hifair™ Ⅲ Reverse Transcriptase, RNase inhibitor, Random Primers/Oligo (dT)18 Primer Mix), RNA template and Rnase-free H2O are added to reverse transcription, and gDNA digester is terminated simultaneously to ensure the integrity of cDNA.

The product is suitable for two-step RT-QPCR detection, specially optimized for qPCR, proportionally optimized Random Primers/Oligo (dT)18 Primer Mix, enabling cDNA synthesis to be initiated from all regions of RNA transcript with the same reverse transcription efficiency. The authenticity and repeatability of qPCR results are guaranteed to the greatest extent. The reverse transcription product is compatible with SYBR Green method and TaqMan probe method qPCR, Hifair™ UNICON™ qPCR SYBR Green Master Mix, Hifair™ qPCR SYBR Green Master Mix or Hifair™ qPCR TaqMan Probe Master Mix can be selected according to experimental purposes, is used in combination to perform high-performance gene expression analysis.


  • High-performance reverse transcription
  • Increase reaction temperature - First strand cDNA can be synthesized in the temperature range of 55 to 60°C
  • Efficient removal of genomic residues
  • Broad linear detection range


  • Construction of cDNA library
  • Antisense RNA synthesis


Type of end product cDNA (First-Strand)
PCR method RT-PCR
Type of reagent Reverse Transcription
Type of sample RNA
Optimal reaction temperature 55℃-60℃
Reverse transcriptase M-MLV


Components No. Name 11141ES10 (10T) 11141ES60 (100T)
11141-A RNase-free H2O 1 mL 2×1 mL
11141-B 5×gDNA digester Mix 30 μL 300 μL
11141-C 4×HifairTM Ⅲ SuperMix plus 50 μL 500 μL

Shipping and Storage

The product is shipped with dry ice and can be stored at -15℃ ~ -25℃ for 18 months.


Figure 1. Excellent reverse transcription efficiency, suitable for genes with different expression abundances with different GC contents

Figure 2. Linear assay range Total RNA 10 pg~5 μg

Figure 3. Efficient removal of genomic residues

Cited from"An RNA-cleaving threose nucleic acid enzyme capable of single point mutation discrimination. Nat Chem . 2022 Mar;14(3):350-359. doi: 10.1038/s41557-021-00847-3."

Cited from"The tetratricopeptide repeat protein OsTPR075 promotes heading by regulating florigen transport in rice. Plant Cell . 2022 Sep 27;34(10):3632-3646. doi: 10.1093/plcell/koac190."

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