Real-Time PCR (qPCR) Guide August 14, 2019 Basically, there are three kinds of real-time PCR (qPCR): ONE-STEP REAL-TIME PCR Use purified RNA. The two reactions of cDNA synthesis by RT (reverse transcription) and real-time PCR happen in one tube. Fewer steps reduce possible contamination. Less pipetting ensures more accurate results. Useful for high-throughput qPCR TWO-STEP REAL-TIME PCR Use purified RNA. Two separate reactions, cDNA synthesis and real-time PCR, in separate tubes. More efficient because random or oligo dT primers can be used. Offers possibility to store cDNA for future use. Can use one cDNA sample for multiple assays with different primers DIRECT REAL-TIME PCR Use purified RNA, or use sample directly, i.e., no purification of RNA is needed. Use sample directly, i.e., no purification of RNA is needed. Reaction system optimized for using cell lysate directly for cDNA synthesis and real-time PCR Cell lysate can be stored for future use. Reduces errors and contamination. Best for rare or unique samples and high-throughput screening. CLICK HERE TO SEE OUR QPCR PRODUCT OPTIONS