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Reverse transcription is a widely used application in molecular biology research for RNA analysis and gene expression studies. This process is carried out by an enzyme called reverse transcriptase, which can convert RNA into a complementary DNA (cDNA) strand. The Thermo Scientific reverse transcriptase portfolio offers robust and reliable reverse transcriptases in a variety of optimized formats for use in RT-PCR and RT-qPCR applications.
Selecting the right reverse transcriptase for cDNA synthesis is critical to detecting low-abundance RNAs and efficiently transcribing RNA molecules with high amounts of secondary structures. When selecting a reverse transcriptase, it is also important to consider factors such as sensitivity and thermostability, which can affect the accuracy and efficiency of the cDNA synthesis. The various formats, include:
Reverse transcriptases are selected for their sensitivity and thermostability. The attributes of various Thermo Scientific reverse transcriptases are listed in Table 1.
Enhanced performance | Routine performance | |||
---|---|---|---|---|
Product | Maxima™ H Minus Reverse Transcriptase | Maxima™ Reverse Transcriptase | RevertAid H Minus Reverse Transcriptase | RevertAid Reverse Transcriptase |
Active temperature range | 50–65 oC | 50–65 oC | 42–55 oC | 42–50 oC |
RNase H activity | No | Yes | No | Yes |
Reaction time | 15–30 min | 15–30 min | 60 min | 60 min |
Sensitivity (total RNA) | 1 pg | 1 pg | 100 pg | 100 pg |
Maximum cDNA length | 20 kb | 20 kb | 13 kb | 13 kb |
Format | Standalone, kit, and master mix | Standalone and kit | Standalone and kit | Standalone and kit |
Primers are an essential component of the reverse transcription reaction, and they are added to the reaction mix along with the reverse transcriptase and other necessary components. The addition of appropriate primers is crucial for successful cDNA synthesis in the reverse transcription reaction. Two commonly used primer types to initiate the synthesis of cDNA are oligo dT primers and random hexamer primers.
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For Research Use Only. Not for use in diagnostic procedures.