Evaluating the Efficiency of Primer Extension Capture as a Method to Enrich DNA Extractions

• NCJ Number: 254109
• Date Published: 2019
• Length: 7 pages
• Author(s): Misa Winters; Cara Monroe; Jodi L. Barta; Brian M. Kemp
• Agencies: NIJ-Sponsored
• Publication Type: Research (Applied/Empirical), Report (Study/Research), Report (Grant Sponsored), Program/Project Description
• Grant Number(s): 2011-DN-BX-K549
• Award: The Behavior of Contamination and Methods for Extraction and Capture of Low Copy Number and Degraded DNA.

Annotation

This study's goal was to document the efficiency of primer extension capture (PEC) as a method to enrich DNA eluates of targeted DNA molecules and remove nontarget molecules from pools containing both.

Abstract

Efficiency of this method was estimated by comparing number of copies in to copies out by quantitative polymerase chain reaction. PEC retention of DNA targets ranging 109©\288 base pairs (bps) in length was 15.88¨C2.14 percent (i.e., loss of 84.12¨C97.86 percent of target molecules). Experimental modifications of the PEC method resulted in no significant improvements; however, the benefit of PEC was revealed in its ability to remove most nontarget DNA molecules (99.99 percent). researchers also found that many (56.69 percent) of the target molecules were lost prior to their immobilization on the streptavidin -coated beads. These estimates of methodological efficiency are directly comparable to previous ones observed following fishing for DNA, an alternative method for DNA enrichment. (publisher abstract modified)