The PCR resource manual.

  • The how-to guide:qPCR from primer design to testing those primers in preparation for quantitative PCR

cDNA synthesis

  • This protocol is used to synthesis cDNA from isolated RNA using the SuperScript II reverse transcriptase enzyme with the aim to use the synthesized cDNA in subsequent 2-stage RT-qPCR reactions.

 Designing primers for qPCR

 Two-step qPCR

Two-step reverse transcription quantitative polymerase chain reaction, often abbreviated as 2-Step RT-qPCR or simply as qPCR, is a high-throughput assay design that allows the real-time measurement of the expression level of specific genes at the RNA level. This protocol was written for SYBR green.