Whole Genome Amplifiction
Recommended Use
If a DNA sample cannot be replaced for any reason, the CGF can perform whole genome amplification (WGA). WGA shouldn't be considered a direct replacement for genomic DNA. Amplified samples can have issues with completion and concordance in genotyping when compared to genomic material. It is to be noted that the quality of the starting material will greatly influence the end product that is produced by WGA. Due to assay constraints, WGA samples cannot be used in Illumina® Infinium®, Custom Infinium® (iSelect)™ or Affymetrix® assays.
Process
The CGF uses the illustra™ GenomiPhi™ series of WGA kits from GE Healthcare. There are two versions in this series. The GenomiPhi™ version 2 kit will produce between 4 and 7 µg of DNA from the starting amount of 50 ng. The GenomiPhi™ HY (high yield) kit will produce between 40 and 50 µg of DNA, also from the 50 ng of starting material. The average product length as reported by GE Healthcare is >10 kb. Once samples have been amplified, the newly amplified sample will be handled according to the CGF Sample Handling pipeline. Read more about the WGA kits.
Requirements
Minimum requirements for samples to be eligible for WGA is at least 50 ng of total DNA at a concentration of 5 ng/µl or greater as determined by our initial concentration measure using the NanoDrop® micro-volume spectrophotometer.
Reporting
A report of samples that did not pass the WGA or sample handling process, and cannot be included in the genotyping project, will be included in the Pre-Genotyping QC report. No formal sample handling report will be delivered, however the status of each sample received will be listed in the sample_status.txt file. See Pre-Genotyping QC Reporting section for details on these files.
