Real-time QPCR Expression Profiling

For relative and absolute quantification of target gene expression

Platform	ABI 7900HT / Roche LightCycler 480 II
Deliverables	Analytical report Raw data Relative quantification using qbase + (BioGazelle) Reference gene stability using GeNorm (BioGazelle)
Research Objectives	For relative or absolute quantification of target genes
Related Services	Expression Profiling Microarrays Digital PCR RNA-seq

Overview

Whether within coding or non-coding regions of a genome, SNPs account for genetic variation within populations that can influence disease development or drug metabolism, to name a few. Life Technologies offer four-and-a-half million predesigned TaqMan® SNP assays that can be used to validate and screen a low to medium number of SNP markers in a high number of samples.

Service Details

Following DNA isolation and quality control, DNA samples are aliquoted onto a 384-well microplate along with the premixed TaqMan assay (probes and primers) and a PCR mastermix. Fluorescence readings are taken pre- and post-cycling using the ABI 7900HT.The normalised fluorescence readings are used to make a genotyping call and the data is displayed in the form of a scatter plot.

Service Deliverables

An analytical report will be provided containing an in-depth analysis of the project results. The report will include allelic discrimination scatterplots as JPEG files and genotype calls in Excel files (raw data). Sample quality control results will also be supplied. Bioinformatics analysis and interpretation of data is optional.

Starting Material

DNA Quantity: 10µl per SNP assay
DNA Concentration: 3-5 ng/µl