The Applied Genomic Technologies Core

TaqMan SNP Genotyping

The AGTC performs Taqman SNP genotyping for both Custom and On Demand Assays on a 96 and 384-well basis.

Instrumentation

• Applied Biosystems 7900HT Sequence Detection System
• Zymark Twister Microplate Handler
• Fixed Position Bar Code Reader

Supported Dyes

• FAM, SYBR, TET, VIC, JOE, NED, NED, TAMRA, ROX

Thermocyclers

• 3 MJ Dyad dual 384-well block
• Applied Biosystems 7900HT Sequence Detection System

Automated Workstations

• Perkin Elmer Janus

Throughput:

• 7,680 samples (one assay) in a 24hr period

Analysis Software:

• ABI Sequence Detection Software V. 2.2.0

Guidelines

Please contact a member of the AGTC staff before you submit a request for service that has never been performed within the Core.

Complete the Sample ID Sheet and Service Request form whenever you submit samples to the AGTC.  Please consult with the AGTC Genotyping staff to determine the best approach for this service submission. 

Sample Submission

• Samples may be submitted using one of the two options below
• When possible, use the same buffer for resuspension/dilution
• An additional charge will be applied if it is necessary to transfer samples to appropriate lab ware.

1. 96-well format: fill columns before rows and assign unique sample names and plate IDs

• Plate = AB-Gene 96-well plate cat # AB- 1100.
• Caps = AB-Gene 8-strip cat # AB- 0602 or 12-strip cat # AB- 0853.
• Sample = 2ng/µl concentration, 4ng per SNP and an additional 40ng (10ul).
• Buffer = T.E. (10mM Tris HCl, 0.1mM EDTA, pH 8.0) or dH2O.

2. 1.5ml Tubes

• Tubes = 1.5ml clear Axygen cat # MTC-150-C.
• Sample = 2ng/µl concentration, 4ng per SNP and an additional 30ng
• Buffer = T.E. (10mM Tris HCl,0.1mM EDTA, pH 8.0) or dH2O.

Reagent Submission

• Primers and probes may be submitted to the core directly, or after receipt from the manufacturer. 
• Follow all handling instructions associated with the reagents. 
• If submitting reagents that have already been used, please indicate the concentration, buffer used to dilute or resuspend, and the storage conditions.

Control Submission

The Core strongly advises the submission of controls for all possible genotypes.  The samples that are submitted by the investigator as controls are used when testing the quality of the assay and without proper controls, it is impossible for the Core to properly determine the assay’s true quality.