High-throughput qPCR using SYBR® Green is a demanding application that requires consistent and reproducible results from low-volume, high-speed assays.
Protocol describes amplification of DNA through quantitative PCR with SYBR Green. Consistent batch-to-batch performance can be achieved with large numbers of PCR reactions.
Optimization of qPCR conditions is important for the development of a robust assay. The two main approaches are optimization of primer concentration and/or annealing temperatures.
Archived Formalin-fixed, Paraffin-embedded (FFPE) tissue samples are invaluable resources for profiling gene expression and studying a variety of diseases.
A protocol that can be used as a basic template for qPCR incorporating SYBR Green I DNA binding dye that is amenable to modification and applicable for use as validation for a set of primers.