Real-Time PCR

Real-Time PCR or Q-PCR has now become an essential component of studies based on quantitation of gene expression, array verification, pathogen detection, viral load estimation and genotyping. In contrast to traditional PCR methods, Real-Time PCR allows measuring the kinetics of the amplification reaction in the initial phases. Traditional methods use time-consuming gel electrophoresis for detection of PCR amplification only at the end of the PCR reaction. Further these methods have limitations such as poor precision, low sensitivity, short dynamic range, low resolution, non-automation etc. In contrast, Real time PCR collects data in the exponential growth phase and quantifies the number of amplicons on the basis of fluorescent signals generated. Further Real Time PCR offers increased dynamic range of detection without any post PCR processing of the amplified samples.

Investigators at the institute are using this facility for quantifying the expression levels of various genes in reproductive tissues/organs under different physiological or pathological conditions. The facility is also being used for SNP genotyping of individuals predisposed to certain reproductive disorders such as congenital adrenal hyperplasia etc. Users bring their Q-PCR reaction samples in a system-compatible (Applied Biosystems-7900HT) 96-well plate format with optical adhesive covers. The system provides data for relative/absolute quantification, melt curve and allele discrimination.

Contact person: Dr. Geetanjali Sachdeva               91-22-24192007.