The Applied Genomic Technologies Core

Multiplex ligation-dependent Probe Amplification (MLPA)

The AGTC optimizes and validates MLPA kits from MRC Holland used to detect gene copy number variation.

DNA Analyzers

• Applied Biosystems 3730 DNA Analyzer (96 capillary instrument)

Supported Dye sets

• (G5): 6-FAM, VIC, NED, PET, LIZ

Thermocyclers

• 6 MJ Dyad dual block
• 4 Techne 96-well

Automated Workstations

• Perkin Elmer Multiprobe II Automated Workstation
• Perkin Elmer Janus
• Perkin Elmer Evolution EP3

Throughput

• 184 samples in a 48hr period

Analysis Software

• Genemarker (SoftGenetics)

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 according the outline below.  When possible, use the same buffer for re-suspension/dilution.  An additional charge will be applied if it is necessary to transfer samples to appropriate lab ware.  Samples must be submitted in 96-well format: fill columns before rows and assign unique sample names and plate IDs

              Plate = Phenix 96-well plate cat # MPC-0296.
              Caps = Phenix 8-strip cat# MPC-440 or 12-strip cat # MPC-660.
              Sample = 5ul at 40ng/µl = total 200ng
              Buffer = T.E. (10mM Tris HCl, 0.1mM EDTA, pH 8.0) or dH2O.

Reagent Submission

MLPA Kits may be submitted to the core directly, or after receipt from MRC Holland.    

Control Submission

The Core strongly advises the submission of five control samples

1.  Three samples that do not contain copy number variation for any probe in the MLPA Kit
2.  One sample with one or more duplications detectable by the MLPA Kit
3.  One sample with one or more known deletions detectable by the MLPA Kit

The control samples above 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.