The Applied Genomic Technologies Core

PCR Cleanup for Sequencing

The AGTC optimizes PCR and uses QIAquick PCR purification columns to purify samples for sequencing.  The core is capable of processing samples on a 96-well basis or individually.  Samples will be quantitated and returned frozen in 1.5ml Eppendorf tubes or 96-well plate.

Instrumentation

• Sigma 4-15C Centrifuge

Thermocyclers

• 6 MJ Dyad dual block
• 4 Techne Genius 

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 re-suspension/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 = Concentration and amount of DNA is assay specific, please consult AGTC Genotyping staff
• 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 = Concentration and amount of DNA is assay specific-consult AGTC Genotyping staff
• Buffer = T.E. (10mM Tris HCl, 0.1mM EDTA, pH 8.0) or dH2O.

Reagent Submission

• Primers may be submitted to the AGTC directly or after receipt from the manufacturer. 
• Follow all handling instructions associated with the reagents. 
• If submitting primers that have already been re-suspended and/or used please indicate the concentration, buffer used to dilute or re-suspend, and the storage conditions (if different from the manufacturers recommendations).

Control Submission

The submission of positive control DNA templates is not necessary if the AGTC has previously carried out the PCR. If requesting clean-up service for a PCR not previously performed by our core, we recommend submitting a control sample for testing. The core will carry out a test PCR using control DNA prior to attempting PCR clean-up. There is an additional charge if the PCR needs to be optimized.