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cDNA Synthesis

Product picture of qPCRBIO cDNA Synthesis Kit

cDNA Synthesis

In molecular biology, complementary DNA (cDNA) is synthesised from an RNA template in a reaction catalysed by the enzyme reverse transcriptase (RTase). cDNA synthesis is the first step in many molecular biology workflows, such as gene expression studies using real-time PCR. cDNA is also used in gene cloning and in the creation of cDNA libraries.

Important considerations when performing cDNA synthesis include the type of primers and the type of RTase used. There are three types of primers used to initiate reverse transcription depending on the RNA template and the downstream application (oligo(dT) primers, random primers, and gene-specific primers). Careful priming is needed to prevent over-representation of cDNA synthesised from the 5′ or 3′ ends of the RNA (5′ or 3′ bias) and to avoid an abundance of truncated cDNA transcripts. PCR Biosystems offers a range of flexible and convenient products for cDNA synthesis, including kits containing premixed random hexamers and anchored oligo(dT) optimised for the generation of cDNA for use in real-time PCR experiments.

The type of RTase used is important as these enzymes can differ in their functional activities and properties. All PCR Biosystems cDNA synthesis products contain a highly active Moloney Murine Leukemia Virus (MMLV) RTase engineered to have a reduced RNase H activity and increased thermostability compared to wild type MMLV, providing higher specificity, higher yields and more full length cDNA product. The ability to perform reactions at higher temperatures can be very helpful when dealing with RNA that contains a high amount of secondary structure.

cDNA Synthesis for qPCR Analysis

Reverse Transcription qPCR (RT-qPCR) is an instrumental technique for the analysis of gene expression. For expression studies to be successful, it is essential for high quality cDNA to be generated…

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Low Copy Number Transcripts

Reverse transcription of RNA to complementary DNA (cDNA) is the first step in many molecular biology workflows including gene expression studies using real-time PCR, gene cloning and in the creation…

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cDNA Cloning

Reverse transcription of RNA to complementary DNA (cDNA) is the first step in many molecular biology workflows including gene expression studies using real-time PCR, gene cloning and in the creation…

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Inhibitor Resistant

Reverse transcription of RNA to complementary DNA (cDNA) is the first step in many molecular biology workflows including gene expression studies using real-time PCR, gene cloning and in the creation…

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High GC

Reverse transcription of RNA to complementary DNA (cDNA) is the first step in many molecular biology workflows including gene expression studies using real-time PCR, gene cloning and in the creation…

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