The real-time PCR (qPCR) assay is an extremely sensitive and reproducible method to measure levels of gene expression in an organism and is widely accepted as a “Gold Standard” for DNA and RNA quantification.
Anewex provides rapid and reliable real time PCR service (qPCR service, quantitative PCR services or qRT-PCR services) for gene expression analysis and gene copy number determination including RNA or DNA extraction, qPCR primer design and validation, qPCR reaction optimization, and complete data analysis including normalization of samples with internal cost controls. Relative and absolute analysis of gene expression by qPCR is available for all known genes in any organisms.
Anewex Experimental plan of real-time PCR services:
– RNA extraction from samples using total RNA isolation kit and/or TRIzol method.
– RNA quantitation and quality assessment using NanoDrop spectrophotometer and/or gel electrophoresis.
– Design qPCR primers using state of the art software and design parameters; Each primer set is validated for specificity by running qPCR and dissociation curve analysis.
– Set up and run the real-time RT-qPCR reactions. High quality total RNA samples are treated using our proprietary gDNA (genomic DNA) Removing Buffer, reverse transcribed to single-strand cDNA, and run in triplicate qPCR alongside no-RT controls.
– Data analysis. The averaged value is normalized to 18S rRNA or reference gene selected by customers for each sample. To evaluate the results a ΔΔCt or ΔΔCq method is used to compare changes in gene expression between controls and treated samples.
We receive:
3 μL cDNA for 1 target gene
Primers
Data Delivery:
Data analysis is performed using our qRT-PCR data analysis software and data can be delivered via email. The primer sequences, raw Ct values, and the analyzed data file including all fold-change information are included in the study report.
Timeline:
* Standard RNA Isolation, RNA quantitation, and Quality Assessment: 2 days
* Reverse Transcription: 1 day
* Primer design, syntheis, and validation: 1 week
* Real-time PCR assay, ΔΔCt or ΔΔCq analysis: 3 days
* Total: 1-2 weeks
