Taqman SNP genotyping assay

eagle-i ID

Resource Type

Properties

1. Fee for service

   Yes

2. Resource Description

   "Taqman Genotyping Assay combines PCR amplification and detection in the same reaction. The method is based on the 5' nuclease activity of Taq DNA polymerase. A PCR is performed using primers that will amplify the DNA region containing the SNP of interest. Included in the reaction are two allele-specific fluorogenic probes, each consisting of a different fluorescent reporter dye and a fluorescent quencher. In the intact probe, the proximity of the quencher to the fluorphore causes fluorescence resonance energy transfer (FRET), reducing the fluorescence from the reporter dye. During PCR, the 5' nuclease activity of Taq digests the allele-specific probe bound to the region of the SNP, releasing the fluorescent dye from the quencher and allowing generation of a fluorescence signal. Depending on which dye signal is generated, the SNP alleles are determined. If only one dye signal is detected, the SNP is homozygous for the allele corresponding to the allele-specific probe, and if both dyes are detected, then the SNP is heterozygous."

3. Service Fee URL

   https://pathbio.med.upenn.edu/pbr/portal/mpf/fees.php

4. Related Resource

   Beckman Coulter BioMek FX robotic workstation

5. Related Resource

   QuantStudio 12K Flex Real-Time PCR System

6. Service Provided by

   Molecular Profiling Facility (Penn)

7. Website(s)

   https://pathbio.med.upenn.edu/pbr/portal/mpf/technology.php#singleassayabiprism

8. Related Technique

   SNP genotyping assay

9. Related Technique

   Real-time PCR