BioReady I Reverse Transcriptase is an engineered version of Moloney Murine Leukemia Virus (M-MLV) reverse transcriptase with reduced RNase H activity and increased thermostability. The enzyme can be used to synthesize higher yield first stranded cDNA at higher temperature than the conventional M-MLV.
1. Low RNase H activity: Reduces RNA degradation during the first-strand cDNA synthesis, resulting in higher yields of full-length cDNA.
2. High thermal stability: The optimal reverse transcription temperature is 42-50 ℃, suitable for RNA templates with complex secondary structures.
3. High purity: No nucleases remain, and host gDNA residue is low.
4. Application range: First-strand cDNA synthesis, one-step RT-PCR.
1. Using extracted total RNA from mouse liver, the total RNA was subjected to a 10-fold gradient dilution and reverse transcribed. One-tenth of the first-strand cDNA product was used as a template for amplification with BSB70 pre-mix, and the expression level of the HBB gene was detected (using a probe-based method). The results are shown in the figure below.
Results: The results show an R² =0.9998, indicating excellent linearity. The reverse transcriptase can effectively reverse transcribe templates at different concentrations, including as low as 10pg of total RNA, demonstrating high amplification efficiency.
2. Using different concentrations of the SARS-CoV-2 RNA as templates, one-step reverse transcription amplification was performed, including the use of BioReady I reverse transcriptase. The expression levels of the N gene, O gene, E gene, and internal reference were detected (using a probe-based method, in quadruplicate). The results of the one-step reverse transcription amplification were compared with similar reagents from an imported manufacturer (T) and a domestic manufacturer (B). The increase in amplification was higher than that of the comparable manufacturer's reagents, and the Ct values were earlier than those of the comparable manufacturer's reagents. The amplification results are shown in the table below:
Results: The results indicate that BioReady I reverse transcriptase has a superior reverse transcription efficiency compared to the reagents from brand T and brand B.
1. The kit should be transported under frozen conditions.
2. Kit storage: Store in a -20°C refrigerator away from light, and the repeated freezing and thawing should not exceed 5 times.
3. Validity period: 12 months, please use within the validity period
