highQu® qScriber™ cDNA Synthesis Kit

cDNA (complementary or copy-DNA) synthesis is a fundamental technique used in both molecular biology research and in diagnostic applications. Reverse transcription process employs reverse transcriptase enzymes (RT), RNase inhibitors, template, random hexamer, oligo dT or specific primers and dNTPs. cDNA is produced aiming to perform different downstream applications. For example, for cloning purposes one takes specific primers, and performs the PCR after the cDNA synthesis to generate long DNA fragments for cloning into the plasmid vectors and for sequencing. For gene expression analysis purposes, short random primers are used, and generated short cDNA fragments unbiased represent the whole genome for qPCR analysis.

A good cDNA synthesis kit provides high yields of high-quality cDNA with minimal bias or variability. For complex targets containing high-GC regions thermostable reverse transcriptases are preferred, as they allow performing the reaction at higher temperature what prevents formation of secondary structures and increases the cDNA yield. Typically, the optimal cDNA synthesis temperature ranges between 37-42°C, depending on the reverse transcriptase enzyme used. Advanced RT enzymes go up to 55°C and above.

Optimizing cDNA synthesis protocol is crucial for generating accurate cDNA libraries. Several factors can influence the quality and yield of cDNA, including the amount and quality of RNA template, the concentration and quality of reverse transcriptase enzyme, quality of the primers, and the cDNA synthesis temperature.

At highQu, we offer several kits and enzymes for cDNA synthesis applications, these include: 1Step RT PCR Kit for fast PCR, HighScriber™ Reverse Transcriptase Mix for cloning purposes, and qScriber™ cDNA Synthesis Kit for a real-time qPCR-based gene expression analysis.