Real-Time PCR Protocols

Protocols and information related to real-time PCR.

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Real-Time PCR Protocols Protocol Links Sort by: Hits | Alphabetical

cDNA Synthesis and Real-Time PCR Using RNA from Laser-Captured Cells Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4108

Protocol describes the real-time PCR using RNA purified from laser-captured tissues Laser Capture Microdissection (LCM): Preparation and Sectioning of Frozen Tissue Blocks and Purification of RNA from Isolated Cells. - [Read cDNA Synthesis and Real-Time PCR Using RNA from Laser-Captured Cells Protocol]

Determination of the CpG Island Methylator Phenotype (CIMP) in colorectal cancer using MethyLight - http://www.natureprotocols.com/2006/06/30/determination_of_the_cpg_islan.php

Protocol provides a detailed, step-by-step protocol of the MethyLight assay for detection of CIMP with high sensitivity and specificity in colorectal cancer using a five marker panel composed of CACNA1G, IGF2, NEUROG1, RUNX3 and SOCS1. - [Read Determination of the CpG Island Methylator Phenotype (CIMP) in colorectal cancer using MethyLight]

Multiplex Real-Time PCR Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4113

In multiplex real-time PCR, different sets of primers with different labels are used to amplify separate genes from the template DNA in one tube. This protocol uses LUX (Light Upon eXtension) primers from invitrogen. FAM (6-carboxy-fluorescein) is used to label the gene of interest, and JOE (6-carboxy-4', 5'-dichloro-2',7'-dimethoxy-fluorescein) is used to label a housekeeping gene as an internal control to normalize between different reactions. - [Read Multiplex Real-Time PCR Protocol]

Real-Time PCR Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4112

Protocol uses FAM-(6-carboxy-fluorescein) or JOE-(6-carboxy-4', 5' -dichloro-2',7' -dimethoxy-fluorescein) labeled LUX (Light Upon eXtension) primers, which can quantify 100 or fewer copies of the target DNA in a background of nonspecific templates, over a broad dynamic range of less than 100-107 copies. It uses uracil deglycosylase (UDG) to minimize the risk of carryover contamination, and includes a melting curve analysis of the product. - [Read Real-Time PCR Protocol]

Real-Time RT-PCR: cDNA Synthesis Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4114

Protocol uses the Superscript II First-Strand Synthesis system for the generation of cDNA from total RNA. RNA purified using TRIzol reagent (Invitrogen) or the methods described in Preparation of RNA Using Guanidinium Isothiocyanate/Cesium Chloride Ultracentrifugation, Preparation of RNA from Paraffin-Embedded Fixed Tissue. - [Read Real-Time RT-PCR: cDNA Synthesis Protocol]

Suppression of Insulin Target Gene Expression by SecinH3 Quantified by Real-Time PCR Protocol - http://www.natureprotocols.com/2006/12/15/suppression_of_insulin_target.php

Protocol for suppression of insulin target gene expression by SecinH3 quantified by real-time PCR. - [Read Suppression of Insulin Target Gene Expression by SecinH3 Quantified by Real-Time PCR Protocol]