AI Article Synopsis
- The study aims to demonstrate that a modified PCR method, known as isolation of PCR products (IPCRp), improves the genotyping of low-copy number DNA in fecal samples compared to traditional methods.
- DNA was extracted from horse feces using the Qiagen DNA Stool Mini Kit, and the IPCRp method amplified the DNA for analysis, significantly reducing background noise during the process.
- The results showed a 100% genotyping success rate, with 87% of samples revealing complete genotype profiles for six loci, indicating that combining the IPCRp method with the extraction kit effectively enhances the recovery of horse DNA genotypes from fecal matter.
Article Abstract
Aim: To show that application of the polymerase chain reaction (PCR) method modified for amplification of a low-copy number DNA samples, ie, the isolation of PCR products (IPCRp), would represent improvement in obtaining genotypes from a fecal DNA compared with previously used genotyping methods.
Methods: 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.
Results: The IPCRp-based multiplex kit (6 loci) revealed equine DNA full genotype profiles, ie, appearance of all six loci, when sampled from fresh feces in 87% of the samples and partial genotype profile (appearance of one to five loci) in 13% of the samples, for a total of 100% genotyping success rate.
Conclusion: 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.
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|---|---|
| http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5470126 | PMC |
| http://dx.doi.org/10.3325/cmj.2017.58.239 | DOI Listing |
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