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Sample Preparation

Sample Preparation

The input DNA amount varies based upon how many wells per sample are required for the specific iPlexPro panel. The standard DNA input amount is 20 ng per well (2 µL per well of a 10 ng/µL DNA solution in water). The Molecular Diagnostics Laboratory will perform sample QC by measuring the DNA concentration using the NanoDrop. If the concentration is below the recommended amount, the investigator will be notified and given the option to proceed with the analysis. The Molecular Diagnostics Laboratory cannot guarantee that the analysis will be successful if the DNA quantity is below the recommended amount.

General Sample Preparation Guidelines

  • Isolate DNA using a commercially available column purification kit (Qiagen, Sigma, etc.) and elute with water
  • Dilute DNA in water to a concentration of 10 ng/µL for iPlexPro
  • Avoid the use of TE or other buffers to minimize the generation of salt adducts
  • DNA should have a 260/280 ratio of 1.8-2.0
  • Please provide a minimum of 15 µL EXTRA per sample for QC and dead volume of our repeater pipettes
  • Only the recommended amount of DNA should be submitted as excess DNA will be discarded
  • DNA can be submitted in microcentrifuge tubes or 96 well plates
  • Submission in 96 well plates is required if your custom assay only uses 1-3 wells/sample (>24 samples to be run on a single plate)
  • Requests should be made using GNomEx and each sample should be labeled according to the submission instructions

UltraSEEK Sample Preparation Guidelines

  • Please follow the general sample preparation guidelines except dilute DNA in water to a concentration of 1 ng/µL
  • Use a fluorescence based DNA quantification method such as Qubit PicoGreen