A Simple Method of Genomic DNA Extraction from Human Samples for PCR-RFLP Analysis
(2013) In Journal of Biomolecular Techniques 24(4). p.224-231- 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... (More)
- 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. (Less)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/a9823322-76ab-4152-939a-43d4c48d8a4b
- author
- Ghatak, Souvik LU ; Muthukumaran, Rajendra Bose and Senthil Kumar, Nachimuthu
- publishing date
- 2013
- type
- Contribution to journal
- publication status
- published
- in
- Journal of Biomolecular Techniques
- volume
- 24
- issue
- 4
- pages
- 224 - 231
- publisher
- Association of Biomolecular Resource Facilities
- external identifiers
-
- scopus:84891553048
- ISSN
- 1524-0215
- DOI
- 10.7171/jbt.13-2404-001
- language
- English
- LU publication?
- no
- id
- a9823322-76ab-4152-939a-43d4c48d8a4b
- date added to LUP
- 2021-11-09 16:27:03
- date last changed
- 2022-04-03 21:34:54
@article{a9823322-76ab-4152-939a-43d4c48d8a4b,
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.}},
author = {{Ghatak, Souvik and Muthukumaran, Rajendra Bose and Senthil Kumar, Nachimuthu}},
issn = {{1524-0215}},
language = {{eng}},
number = {{4}},
pages = {{224--231}},
publisher = {{Association of Biomolecular Resource Facilities}},
series = {{Journal of Biomolecular Techniques}},
title = {{A Simple Method of Genomic DNA Extraction from Human Samples for PCR-RFLP Analysis}},
url = {{http://dx.doi.org/10.7171/jbt.13-2404-001}},
doi = {{10.7171/jbt.13-2404-001}},
volume = {{24}},
year = {{2013}},
}