A Simple Method of Genomic DNA Extraction from Human Samples for PCR-RFLP Analysis

Abstract

Isolation of DNA from blood and buccal swabs in adequate quantities is an integral part of forensic research and analysis. The present study was performed to determine the quality and the quantity of DNA extracted from four commonly available samples and to estimate the time duration of the ensuing PCR amplification. Here, we demonstrate that hair and urine samples can also become an alternate source for reliably obtaining a small quantity of PCR-ready DNA. We developed a rapid, cost-effective, and noninvasive method of sample collection and simple DNA extraction from buccal swabs, urine, and hair using the phenol-chloroform method. Buccal samples were subjected to DNA extraction, immediately or after refrigeration (4 -6°C) for 3 days. The purity and the concentration of the extracted DNA were determined spectrophotometerically, and the adequacy of DNA extracts for the PCR-based assay was assessed by amplifying a 1030-bp region of the mitochondrial D-loop. Although DNA from all the samples was suitable for PCR, the blood and hair samples provided a good quality DNA for restriction analysis of the PCR product compared with the buccal swab and urine samples. In the present study, hair samples proved to be a good source of genomic DNA for PCR-based methods. Hence, DNA of hair samples can also be used for the genomic disorder analysis in addition to the forensic analysis as a result of the ease of sample collection in a noninvasive manner, lower sample volume requirements, and good storage capability.

Comparison between Two Different DNA Extraction Techniques Taken from Buccal Swabs Suitable for Genetic Analyzer

“…Organic extraction method worked well for recovery of high molecular weight DNA. This finding is consistent with what have been observed in previous studies [16,21], while the highest purity was noticed in the kit. This type of kits was made for isolation DNA from trace sample (forensic casework) and was manufactured to release any inhibiters with high quality.…”

Extracellular matrix protein 1 gene rs3737240 single nucleotide polymorphism is associated with ulcerative colitis in Turkish patients

“…Afterward, genomic DNA extraction was performed with the salting-out method (17). Buccal mucosa samples were collected from the healthy control group and genomic DNA extraction was performed with the standard phenol/chloroform method (18). …”

Mutation Profiling in Mitochondrial D-Loop Associated with Stomach Cancer and Tobacco Consumers

“…DNA was extracted from the samples by following the modified protocol of Ghatak et al (2013) [13]. The extracted DNA was dissolved in 1XTE buffer (pH 8.0) and stored at -20ºC until used.…”