Do you feel that it is too cumbersome to do two steps of reverse transcription?
Are you concerned that double pipetting increases the risk of RNA degradation?
Then you must try this HiScript® III All-in-one RT SuperMix Perfect for qPCR (Vazyme #R333, click to see the detail), which takes only 15 min to complete Genomic DNA (gDNA) cleanup + Reverse Transcription in one step!
HiScript® III All-in-one RT SuperMix Perfect for qPCR (Vazyme #R333) (Figure 1), as the name suggests, ''All-in-one'' is a one-tube reaction that can effectively reduce the risk of sample contamination and RNA degradation during the Operation process.
Figure 1: Physical view of R333
This product contains heat-sensitive DNase, HiScript III Reverse Transcription, and the optimal buffer optimized for the one-step reaction of gDNA removal and reverse transcription, which can eliminate the residual gDNA in the RNA template while completing the reverse transcription reaction.
Ultra-high cDNA stability: Heat-sensitive DNase, more thorough inactivation, and more stable cDNA storage for a long time.
Simple and fast operation: gDNA cleanup and reverse transcription are completed in one step. Just add RNA template, and the reaction can be completed in 15 min (Figure 2).
Figure 2: Workflow of Reverse Transcription
Excellent Reverse Transcription efficiency
Using 10 ng / 1 μg RNA of Hela cells, Mouse liver, and Arabidopsis leaves as templates. Vazyme # R333, competing All-in-one products (Supplier TG-H, TG-E, TI) and common products (Supplier Vazyme #R323, TA) were used for reverse transcription, respectively. The obtained cDNA was quantitatively analyzed by qPCR.
It can be seen from the figure 3 that Vazyme #R333 has the smallest amplification CT value on different templates, indicating that Vazyme #R333 has better reverse transcription efficiency.
Figure 3: The CT values of different experimental protocols
Ultra-high cDNA stability
Reverse transcription was carried out with 100 ng of RNA (HeLa cell) through Vazyme #R333. The obtained cDNA was stored at 37°C for 7 days, 4°C for 4 weeks, and freeze-thawed 30 times for testing, while the cDNA was stored at -20°C as a control.
It can be seen from the figure 4 that the ΔCT of cDNA quantitatively obtained under various conditions is within the range of 0.5 (ΔCT is within ±0.5, and the difference between the two is considered to be negligible), indicating that the cDNA obtained by reverse transcription of Vazyme #R333 can be Stored at 37°C for 7 days, 4°C for 4 weeks, and repeated freezing and thawing 30 times, the cDNA still has good stability.
Figure 4: The ΔCT values of cDNA under different storage conditions