Quantitative Real-Time PCR Core

Overview

This Core offers quantitative measurement of nucleic acid (DNA or RNA) levels using real-time PCR.

This includes equipment and software for transcript quantitation, gene expression analysis, PCR melting curves, SNP genotyping, whole-genome analysis, viral particle titration. The heart of the Core is the Applied Biosystems 7500 and 7000 real-time PCR systems with software for primer selection and data analysis.

Real-time quantitative PCR provides improved sensitivity, accuracy, speed, and throughput over standard PCR methods. Detection is based on TaqMan fluorescent probes with 5' reporter and 3' quencher labels, such that synthesis from flanking primers leads to cleavage of the 5' reporter dye and detectable fluorescence.

TaqMan probes can be synthesized with different color dyes permitting the analysis of multiple sequences in a single PCR reaction, so that each reaction can contain an internal control. The TaqMan probes can also be used for allelic discrimination assays. SYBR green can be used in place of the TaqMan probes to detect total PCR product.

Getting Started

To gain access to the facility and to book a time slot, follow these steps

1. Go to http://researchcores.partners.org
2. Create an account to access partners research cores (if you wish to use your partners password, do not enter any password and your partners password will be set as default).
3. To add a fund #, select User, Fund, add new fund #, Assign Fund.
4. Select Cores, Real-Time PCR, MGH Quantitative Real-Time PCR, Schedule, Select Machine under equipment Menu and Schedule Time.

Once you have done this, contact our technician (mghvectorcore@partners.org) and he or she can grant you access to the facility.