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qPCR Gene Expression/Copy Number Analysis Protocol Using SYBR Green I Dye Detection
Measuring a target quantity relative to one or more stable reference genes using SYBR Green I dye detection is a common application of qPCR. Below is a standard protocol that can be adapted to specific experimental needs.
Primer Optimization Protocol Using Temperature Gradient
Gradient PCR for assay optimization is to determine the optimum annealing temperature (Ta) of the primers by testing identical reactions containing a fixed primer concentration, across a range of annealing temperatures.
KiCqStart™ Universal SYBR® Green qPCR Protocol
Quantitative PCR protocol using SYBR Green reagents. Procedure supports most qPCR instruments.
qPCR Efficiency Determination Protocol
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.
qPCR Reference Gene Selection Protocol
Analysis of gene expression data requires a stable reference or loading control. This reference is usually one or more reference genes.
Primer Concentration Optimization Protocol
Primer Concentration Optimization Protocol is an approach to create a matrix of reactions. This is used to test a range of concentrations for each primer against different concentrations of the partner primer.
SYBR® Green I Dye Quantitative PCR Protocol
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.
SPUD Assay for Detection of Assay Inhibitors Protocol
he SPUD assay is one option for identification of inhibitors that may be present in RNA or DNA samples. The assay is particularly useful when a large number of samples are to be analyzed or when targets are present at...
Polymerase Chain Reaction
The polymerase chain reaction is one of the most widely used techniques in molecular biology. The PCR process consists of three main steps, Denaturation, Annealing & Extension
RT-qPCR – Quantitative Reverse Transcription PCR
RT-qPCR, or quantitative reverse transcription PCR, combines the effects of reverse transcription and quantitative PCR or real-time PCR to amplify and detect specific targets. RT-qPCR has a variety of applications including quantifying gene expression levels, validating RNA interference (RNAi), ...
Quantitative PCR Basics
Real-time polymerase chain reaction allows researchers to estimate the quantity of starting material in a sample. It has a much wider dynamic range of analysis than conventional PCR
Universal SYBR Green qPCR Protocol
Our SYBR Green qPCR Protocol is a method designed to detect accurate quantification of gene expression and RT-PCR reactions
PCR Assay Optimization and Validation
PCR assay guide navigates you through primer validation and other assay optimization factors to ensure high sensitivity and specificity for optimum DNA/ RNA quantification.