Open Access
A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity
Andrew P. Feinberg
- Vol. 137, pp 266-267
TLDR
In this article, a technique for conveniently radiolabeling DNA restriction endonuclease fragments to high specific activity is described, where DNA fragments are purified from agarose gels directly by ethanol precipitation and are then denatured and labeled with the large fragment of DNA polymerase I, using random oligonucleotides as primers.Abstract:
A technique for conveniently radiolabeling DNA restriction endonuclease fragments to high specific activity is described. DNA fragments are purified from agarose gels directly by ethanol precipitation and are then denatured and labeled with the large fragment of DNA polymerase I, using random oligonucleotides as primers. Over 70% of the precursor triphosphate is routinely incorporated into complementary DNA, and specific activities of over 10(9) dpm/microgram of DNA can be obtained using relatively small amounts of precursor. These "oligolabeled" DNA fragments serve as efficient probes in filter hybridization experiments.read more
Citations
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The C. elegans heterochronic gene lin-4 encodes small RNAs with antisense complementarity to lin-14
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A novel potent vasoconstrictor peptide produced by vascular endothelial cells.
Masashi Yanagisawa,Hiroki Kurihara,Hiroki Kurihara,Sadao Kimura,Yoko Tomobe,Mieko Kobayashi,Youji Mitsui,Yoshio Yazaki,Katsutoshi Goto,Tomoh Masaki +9 more
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WAF1, a potential mediator of p53 tumor suppression
Wafik S. El-Deiry,Takashi Tokino,Victor E. Velculescu,Daniel B. Levy,Ramon Parsons,Jeffrey M. Trent,D Lin,W. Edward Mercer,Kenneth W. Kinzler,Bert Vogelstein +9 more
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A system of shuttle vectors and yeast host strains designed for efficient manipulation of DNA in Saccharomyces cerevisiae.
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Identification of the cystic fibrosis gene: cloning and characterization of complementary DNA.
John R. Riordan,Johanna M. Rommens,Batsheva Kerem,Noa Alon,Richard Rozmahel,Zbyszko Grzelczak,Julian Zielenski,Si Lok,Natasa Plavsic,Jia Ling Chou,Mitchell L. Drumm,Michael C. Iannuzzi,Francis S. Collins,Lap-Chee Tsui +13 more
TL;DR: A deletion of three base pairs that results in the omission of a phenylalanine residue at the center of the first predicted nucleotide-binding domain was detected in CF patients.
References
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Book
Molecular Cloning: A Laboratory Manual
TL;DR: Molecular Cloning has served as the foundation of technical expertise in labs worldwide for 30 years as mentioned in this paper and has been so popular, or so influential, that no other manual has been more widely used and influential.
Journal ArticleDOI
Detection of specific sequences among DNA fragments separated by gel electrophoresis.
TL;DR: This paper describes a method of transferring fragments of DNA from agarose gels to cellulose nitrate filters that can be hybridized to radioactive RNA and hybrids detected by radioautography or fluorography.
Arapid alkaline extraction procedure forscreening recombinant plasmid DNA
TL;DR: The method is simple enough to permit the analysis by gel electrophoresis of 100 or more clones per day yet yields plasmid DNA which is pure enough to be digestible by restriction enzymes, and achievesequate pH control without using a pH meter.
Journal ArticleDOI
Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I
TL;DR: Labeled DNAs (and restriction endonuclease fragments derived from them) are useful probes for detecting rare homologous sequences by in situ hybridization and reassociation kinetic analysis.
Journal ArticleDOI
Screening lambdagt recombinant clones by hybridization to single plaques in situ
WD Benton,Ronald W. Davis +1 more
TL;DR: A rapid, direct method for screening single plaques of Agt recombinant phage is described, which allows at least 10(6) clones to be screened per day and simplifies physical containment of recombinants.