The measure of quality in DNA sample processing starts with an effective nucleic acid isolation procedure. Most problems with DNA sample typing can be attributed to low quantity DNA and/or to the presence of inhibitors in the sample; therefore, establishing which isolation method is best at removing potential inhibitors may help overcome some of the problems analysts encounter by providing useful information in the determination of the optimal approach for any given sample. The methods investigated in the current study included both liquid/liquid (phenol-chloroform) and solid phase based robotic procedures, (PrepFiler and EZ1 chemistries). Following extraction, samples were analyzed by DART in order to determine the level of remaining inhibitors and then quantified and amplified to determine the effect any remaining inhibitor had on the overall results. The data suggest that organic extraction methods result in detrimental amounts of phenol carryover, and automated methods may produce carry‐over of bile salts and other chemicals that preferentially bind the solid phase matrix. Both of these effects can have a negative impact in downstream sample processing and genotyping by PCR. (Publisher abstract modified)
The Use of Direct Analysis in Real Time (DART) To Assess the Levels of Inhibitors Co-extracted With DNA and the Associated Impact in Quantification and Amplification
NCJ Number
252340
Journal
Electrophoresis Volume: 37 Issue: 2 Dated: 2016 Pages: 2807-2816
Date Published
March 2016
Length
10 pages
Annotation
This study used direct analysis in real time (DART) mass spectrometry to investigate the ability of different extraction methods to remove PCR inhibitors.
Abstract
Date Published: March 1, 2016