Journal ArticleDOI
The use of biotinylated DNA probes in parentage testing: non-isotopic labeling and non-toxic extraction.
TLDR
By manipulating probe sizes, blocking agents, selection of membrane and detection system, it is feasible to use non‐isotopic labeling and detection in routine parentage testing and Reproducible results were obtained.Abstract:
With a few exceptions DNA probing techniques require the use of radioisotopes and toxic DNA extraction techniques which render the method expensive, potentially hazardous and time-consuming. Most isotopic labeling techniques use the isotope 32P and require 3-10 days to visualize bands after hybridization. An alternative approach is based on the use of non-isotopic detection methods. The available non-isotopic techniques were assessed and their practicality tested. All probes analyzed were tested on samples extracted with a non-toxic extraction procedure using 6 M NaCl as the substitute for phenol and isochloroform. By manipulating probe sizes, blocking agents, selection of membrane and detection system, it is feasible to use non-isotopic labeling and detection in routine parentage testing. Reproducible results were obtained with labeling a variety of DNA probes of various sizes, plasmid and inserts. With an absence of waste disposal costs, probes that are stable for over two years and a staining procedure which takes 3-5 h versus days the technique is well suited for a normal laboratory setting. The next key to the acceptability of DNA testing will be the commercial availability of DNA probes for widespread use.read more
Citations
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Journal ArticleDOI
Chelex 100 as a Medium for Simple Extraction of DNA for PCR-Based Typing from Forensic Material
Walsh Ps,Metzger Da,Higuchi R +2 more
TL;DR: The extraction of DNA from semen and very small bloodstains using Chelex 100 is as efficient or more efficient than using proteinase K and phenol-chloroform extraction and DNA extracted from bloodstain seems less prone to contain PCR inhibitors when prepared by this method.
Journal ArticleDOI
Simple procedure of DNA isolation from human serum
Journal ArticleDOI
Optimization of the PCR for Detection of Staphylococcus aureus nuc Gene in Bovine Milk
Chang-Hyun Kim,Muhammad Jawad Khan,Dawn E. Morin,Walter L. Hurley,D.N. Tripathy,Marcus E. Kehrli,A.O. Oluoch,Ibulaimu Kakoma +7 more
TL;DR: It is concluded that Tth-PCR on milk samples with the purification of crude DNA extracts using Chelex-100 is as sensitive as but faster than conventional milk bacteriological culture techniques and is highly specific.
Journal ArticleDOI
High prevalence of mutations at codon 249 of the p53 gene in hepatocellular carcinomas from Senegal.
TL;DR: There is an association between countries of high aflatoxin intake and a high frequency of mutation in codon 249 of p53 gene, and that HBV alone does not contribute to these base changes.
Book
Advances in Mutagenesis Research
TL;DR: Advances in mutagenesis research 5, Advances in Mutagenesis Research 5 , مرکز فناوری اطلاعات و اصاع رسانی, کسورزی.
References
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Journal ArticleDOI
A simple salting out procedure for extracting DNA from human nucleated cells
TL;DR: A rapid, safe and inexpensive method was developed to simplify the deprotein-ization procedure that yielded quantities comparable to those obtained from phenol-chloroform extractions, rendering the entire process of RFLP analysis free of toxic materials.
Journal ArticleDOI
Enzymatic synthesis of biotin-labeled polynucleotides: novel nucleic acid affinity probes
TL;DR: Biotin-labeled polynucleotides, both single and double-stranded, are selectively and quantitatively retained on avidin-Sepharose, even after extensive washing with 8 M urea, 6 M guanidine hydrochloride, or 99% formamide.
Journal ArticleDOI
Rapid and sensitive colorimetric method for visualizing biotin-labeled DNA probes hybridized to DNA or RNA immobilized on nitrocellulose: Bio-blots.
TL;DR: At high probe concentrations (250--7560 ng/ml), biotin-labeled DNA exhibits lower nonspecific binding to nitrocellulose than does radiolabeled DNA, so hybridization times required for the analysis of unique mammalian gene sequences can be decreased to 1--2 hr.
Journal ArticleDOI
A genetic linkage map of the human genome
Helen Donis-Keller,Philip Green,Cynthia Helms,Samuel Cartinhour,Barbara Weiffenbach,Karen Stephens,Tim Keith,Donald W. Bowden,Douglas Smith,Eric S. Lander,David Botstein,Gita Akots,Kenneth S. Rediker,Thomas C. Gravius,Valerie A. Brown,Marcia Rising,Carol A. Parker,Jody A. Powers,Diane E. Watt,Erick R. Kauffman,Angela Bricker,Pamela Phipps,Hans Müller-Kahle,Thomas R. Fulton,Siu Ng,James W. Schumm,Jeffrey C. Braman,Robert G. Knowlton,D. Barker,Steven M. Crooks,Steven E. Lincoln,Mark J. Daly,Jeff Abrahamson +32 more
TL;DR: This work reports the construction of a linkage map of the human genome, based on the pattern of inheritance of 403 polymorphic loci, including 393 RFLPs, in a panel of DNAs from 21 three-generation families, by a combination of mathematical linkage analysis and physical localization of selected clones.
Journal ArticleDOI
Non-radioactive hybridization probes prepared by the chemical labelling of DNA and RNA with a novel reagent, photobiotin
TL;DR: The sensitivity of detection of target RNA in dot-blots and Northern blots was equivalent to that obtained with 32p-labelled DNA probes, and photobiotin was also used for the labelling of proteins with biotin.
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