qPCR
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KAPA SYBR® FAST One-Step qRT-PCR Kits FAQs
Frequently asked questions (FAQs) for KAPA SYBR® FAST One-Step qRT-PCR Kits.
RT-qPCR Basic Troubleshooting
Learn the basics of real-time PCR and qPCR with tips, tricks and general troubleshooting.
SYBR® Green Extract-N-Amp™ Tissue PCR Kit Protocol
The SYBR® Green Extract-N-Amp™ Tissue PCR Kit contains all the reagents needed for rapid extraction, amplification and detection of genomic DNA from mouse tails and other animal tissues, buccal swabs, hair shafts, and saliva.
ChIP-qPCR and Data Analysis
Chromatin Immunoprecipitation quantitative real-time PCR (ChIP-qPCR) is commonly used in studies that focus on specific genes and potential regulatory regions across differing experimental conditions and data analysis. qPCR enables DNA analysis in real time by analyzing fluorescent signal intensities that
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
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.
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.
Multiplex qPCR Protocol
A protocol that can be used as a basic template for qPCR incorporating up to four detection probes. In these reactions, primers and probe are included at a final concentration of 200 nM and are run using LuminoCt ReadyMix.
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.
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.
qPCR Using a Single Detection Probe Protocol
A protocol that can be used as a basic template for qPCR incorporating a detection probe that is specific to a single target. In these reactions, primers and probe are included at a final concentration of 200 nM and are
KiCqStart™ One-Step Probe RT-qPCR ReadyMix™
KiCqStart One-Step Probe RT-qPCR ReadyMix is a ready-to-use, highly sensitive master mix for reverse transcription quantitative PCR (RT-qPCR) of RNA templates using hybridization probe detection chemistries such as TaqMan 5'-hydrolysis probes on real-time PCR systems.
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
The 3'/5' Assay for Analysis of RNA Integrity Protocol
The 3’/5’ integrity assay is a potential first step in the identification of RNA degradation. The assay is particularly useful when a large number of samples are to be analyzed or when the degradation is less than that detected by
High-throughput Real-Time PCR: SYBR® Green
High-throughput qPCR using SYBR® Green is a demanding application that requires consistent and reproducible results from low-volume, high-speed assays.
FFPE Tissue
Archived Formalin-fixed, Paraffin-embedded (FFPE) tissue samples are invaluable resources for profiling gene expression and studying a variety of diseases.
dPCR Protocol
The Bio-Rad digital droplet system is based on oil emulsification technology and uses a standard 96-well plate format. Each sample is partitioned into 20,000 reactions, yielding a total of 1.9 million reactions per plate.
MystiCq® MicroRNA® Quantitation System
Method for purification, reverse transcription and quantitative PCR for MicroRNAs using Mysticq reagents
Multiplex Real-Time PCR
Multiplex qPCR employing probe-based chemistries is a demanding application that often requires extensive optimization and validation.
Rapid SYBR® Green qPCR on the Illumina Eco™ Real-Time PCR System
Fast real-time PCR is a demanding application that requires consistent and reproducible results from the difficult amplicons and low-volume reactions
SYBR® Green JumpStart™ Taq ReadyMix™
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.
RNA Immunoprecipitation qPCR (RIP- qPCR) Protocol
RNA immunoprecipitation (RIP) can identify specific RNA molecules of many types associated with specific nuclear or cytoplasmic binding proteins.
Protocol for Conjugating NHS-Ester Modifications to Amino-Labeled Oligonucleotides
Protocol for conjugating NHS-ester modifications to an oligonucleotide with an amino label.
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.
Reverse Transcription Protocol (One-step Probe Detection)
Reverse transcription (RT) is the process of converting RNA to cDNA using a reverse transcriptase enzyme and dNTPs.
KiCqStart™ Universal SYBR® Green qPCR Protocol
Quantitative PCR protocol using SYBR Green reagents. Procedure supports most qPCR instruments.
KiCqStart™ Probe qPCR ReadyMix™
KiCqStart™ Probe qPCR ReadyMix™ is an advanced qPCR reagent system for both fast and conventional PCR cycling protocols or instruments.
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.