Accurate diagnosis starts with reverse transcription

Accurate diagnosis starts with reverse transcription

- RT-qPCR powered by highly efficient reverse transcriptase

Reverse Transcriptase (RT enzyme) is an enzyme capable of converting RNA templates into complementary DNA (cDNA). In molecular biology research, reverse transcriptase is widely used, especially playing an indispensable role in the field of molecular diagnostics. Through the process of reverse transcription, the RNA genome of viruses can be converted into cDNA, which can then be amplified and detected using techniques such as qPCR. This characteristic enables accurate diagnosis of RNA viruses, such as HIV, influenza A, and influenza B.

Figure 1. Schematic diagram of reverse transcription process

The pursuit of RT-qPCR molecular diagnostic reagents that are faster, more sensitive, and easier to operate has become the current major trend of development. As one of the key raw materials in the RT-qPCR process, reverse transcriptase needs to possess faster reverse transcription speed, higher synthesis yield, and better heat resistance. In order to obtain a reverse transcriptase more suitable for RT-qPCR, Yeasen has developed the HifairTM series of reverse transcriptases through genetic modification based on M-MLV, featuring lower RNase H activity, higher thermal stability, and stronger continuous synthesis capability.

Figure 2. Schematic of reverse transcription action

Hifair™Reverse Transcriptases

Yeasen has a variety of HifairTM reverse transcriptase, and can provide a variety of product forms, including enzyme and premix, you can choose according to actual needs. The following table lists the characteristics and recommended applications of different Yeasen reverse transcriptases.

Table 1 The selection guidelines of Hifair™ Reverse Transcriptases

Product Name (Product Number )

Product features

Recommended application

HifairTM III reverse transcriptase (11111ES)

Thermally stable (up to 65°C), suitable for complex templates

RT-LAMP

HifairTM V reverse

transcriptase (11300ES)

Strong compatibility, fast speed, and high synthesis yield

One-step RT-PCR/qPCR

HifairTM Ultra Reverse Transcriptase (14604ES)

Strong continuous synthesis capability, with a synthesis length up to 20 kb

RT-PCR/qPCR, RNA-seq, single cell sequencing

Note: All the enzymes mentioned above are available in high-concentration or non-glycerol formulations.

We now highlight a reverse transcriptase used in RT-qPCR molecular diagnostic reagents: HifairTM V Reverse Transcriptase. This enzyme is distinguished by its strong compatibility, rapid speed, and high synthesis yield, capable of enduring reaction temperatures up to 58°C. It is particularly well-suited for the development of RT-qPCR molecular diagnostic reagents.

Performance demonstration of Hifair™ V reverse transcriptase

1.Capable of withstanding a reaction temperature of 58°C, it is suitable for the reverse transcription of complex RNA templates.

    Figure 3. The Reverse Transcriptase of HifairTM V Reverse Transcriptase and the brand T* were incubated at 55℃ and 58℃ respectively for 30min to detect the residual enzyme activity. The results showed that HifairTM V Reverse Transcriptase could still maintain 70% activity after incubation at 58℃ for 30min, which was much higher than that of the brand T*.

    2.The detection rate of RT-qPCR was better than that of other brands,and the detection range was 1 pg-1 μg.

    Figure 4. Using 1 pg-1 μg (7 gradients) total RNA of 293T cells as template, reverse transcription was performed using HifairTM V one-step RT-gDNA digestion SuperMix for qPCR(Cat#11142), V*, T*. The obtained cDNA was quantitatively analyzed by qPCR. The results showed that the sensitivity of 11142ES could reach 1 pg, which was better than that of other brands.

    The reverse transcriptase mutant Library of Yeasen

    Yeasen continues to promote the evolution of reverse transcriptase with its ZymeEditorTM platform. So far, we have screened out a variety of reverse transcriptase mutants with excellent performance in different application, which can fully meet the needs of different customer groups in diversified application fields.

    Table 2. The reverse transcriptase Mutant Selection Guide of Yeasen

    Recommended Products

    Product Classification

    Product Name

    Catalog No.

    High performance PCR enzyme series

    Hieff UNICON™ Hotstart E-Taq DNA Polymerase, 5 U/μL

    10726ES

    Hifair™ V Reverse Transcriptase(200 U/μL)

    11300ES

    Uracil DNA Glycosylase (UDG/UNG), 1 U/μL

    14455ES

    Murine RNase Inhibitor(40 U/µL)

    10603ES

    dNTP Mix(25 mM each)

    10125ES

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