Genotyping Horse Epithelial Cells From Fecal Matter by Isolation of Polymerase Chain Reaction Products

• NCJ Number: 252098
• Journal: Croatian Medical Journal Volume: 58 Issue: 3 Dated: 2017 Pages: 239-249
• Date Published: 2017
• Author(s): Pero Dimsoski
• Length: 11 pages
• Grant Number(s): 2011-NE-BX-K550
• Agencies: NIJ-Sponsored
• Publication Type: Research (Applied/Empirical); Report (Study/Research); Report (Grant Sponsored); Program/Project Description

Annotation

This article presents the findings and methodology of a project whose goal was to show that application of the polymerase chain reaction (PCR) method modified for amplification of a low copy number DNA samples, i.e., the isolation of PCR products (IPCRp), would represent improvement in obtaining genotypes from a fecal horse DNA compared with previously used genotyping methods.

Abstract

The DNA from the horse fecal matter was extracted by modified Qiagen DNA Stool Mini Kit protocol. Following the extraction, the DNA genotypes from fecal samples were obtained by the most powerful PCR amplification method, the IPCRp. The IPCRp-based multiplex kit amplified biotin-labeled strands were captured on streptavidin-coated plates, where everything but the dye-labeled target sequence was washed, eliminating all the background noise, released, and run on a genotyping instrument in a single-strand configuration. The IPCRp-based multiplex kit (6 loci) revealed equine DNA full genotype profiles, i.e., appearance of all six loci, when sampled from fresh feces in 87 percent of the samples and partial genotype profile (appearance of one to five loci) in 13 percent of the samples, for a total of 100 percent genotyping success rate. The project concluded that these results indicate that the IPCRp amplification method, coupled with the Qiagen DNA Stool Mini Kit extraction can maximize the likelihood of obtaining horse DNA genotypes from fecal samples. (Publisher abstract modified)