http://biowww.net/ Fri, 04 Jul 2008 12:31:54 +0100 FeedCreator 1.7.2 http://biowww.net/detail-795.html A basic protocol of Analysis of DNA by Restriction Enzyme Cleavage and Agarose Gel Electrophoresis for beginners. (David Ng) ... biowww http://biowww.net/detail-925.html Protocol by Patricia V. King, Robert W. at Blakesley Bethesda Research Laboratories. (Marimoto lab at Northwestern University). References: 1. Engler, M.J. and Richardson, C.C. (1982) p. 3. The Enzymes. Vol. XV. Academic Press, New York. 2. Phei ... biowww http://biowww.net/detail-1043.html Protocols of restriciton digestion from Donis-Kellor lab manual. Restriction Enzyme Digests Restriction Enzyme Buffers Restriction Digestion of Plasmid, Cosmid, and Phage DNAs Manual Method of Restriction Digestion of Human DNA Restriction Dige ... biowww http://biowww.net/detail-1110.html A nice troubleshooting guide and FAQs about Restriction Endonucleases from Fermentas. Content: Problems: Undigested or incompletely digested DNA Additional bands atypical for the banding pattern of a particular substrate Diffused DNA zone ... biowww http://biowww.net/detail-2.html Double restriction digestion of DNA involves simultaneously DNA double digestion using two restriction enzymes. It is important to chose right enzymes with compatible buffers so that both enzymes have high activity. Most vendors provide restriction digestion b ... biowww http://biowww.net/detail-19.html I start out with high molecular weight DNA. When I try and carryout my digestion it seems as though my DNA is being degraded by the enzyme and I have tried two different enzymes. When I just add water or buffer or water/buffer to the DNA it is fine i.e. high m ... biowww http://biowww.net/detail-109.html I am genotyping the mice for defective leptin receptor allele (db). According to the protocol I have to run Rsa digestion on the PCR products in order to digest 135 bp fragments into 108 and 25 bp pieces. It seems to work for heterozygotes (I get two bands), b ... biowww http://biowww.net/detail-113.html I had to label about 40-50 primers, and while doing this I have spotted a strange phenomenon: the labelling efficiency could drop to less then 10% depending on the first nucleotide of the primer, independently of the sequence and condition. ... ... ... biowww http://biowww.net/detail-139.html I would like to limit the use of 32P in my lab and I am shopping for non-radioactive labelling methods (random prime). I am primarily interested a system that is successful with Northern blot Hybridizations. I would like to know if anyone would recommend a par ... biowww http://biowww.net/detail-140.html I am trying to digest a plasmid with BclI whose optimum temperature is at 50C. With time, water evaporates and obviously affects the reaction environment. I am aware of some simple tricks like using mineral oil, but am not certain. Anyone ... ... ... biowww http://biowww.net/detail-147.html my collegue does not denature the DNA template before random primer labelling and claims to get efficient incorporation of 32P-dCTP. Is denaturation of the template a "MUST"? I always do , nevertheless. ... ... ... biowww http://biowww.net/detail-207.html I have performed a digest on a vector (pCMV) to use in a ligation. How can I tell if the digest was successful? The difference in size of the undigested and digested vector is only 30 base pairs. I have not run on a gel because the undigested is 4.3 kb and I d ... biowww http://biowww.net/detail-1111.html It is not uncommon to have difficulties in digesting DNA with restriction enzymes. At times, the DNA does not appear to cut at all and sometimes it cuts only partially. If the sequence is known, restriction sites can be predicted with accuracy, but in the lab, ... biowww http://biowww.net/detail-686.html The Restriction Enzyme data BASE is a collection of information about restriction enzymes and related proteins. It contains published and unpublished references, recognition and cleavage sites, isoschizomers, commercial availability, methylation sensitivity, c ... biowww http://biowww.net/detail-1091.html IntEnz is the name for the Integrated relational Enzyme database and is the most up-to-date version of the Enzyme Nomenclature. The Enzyme Nomenclature comprises the recommendations of the Nomenclature Committee of the International Union of Biochemistry and M ... biowww http://biowww.net/detail-687.html RestrictionMapper is a web site that finds restriction endonuclease cleavage sites in DNA sequences. It supports linear and circular DNA and provides several ways to sort and filter output. Also provided is a virtual digest function that simulates a simultaneo ... biowww http://biowww.net/detail-793.html This tool will take a DNA sequence and find the large, non-overlapping open reading frames using the E.coli genetic code and the sites for all Type II restriction enzymes that cut the sequence just once. By default, only enzymes available from NEB are used, bu ... biowww http://biowww.net/detail-794.html This tools perform a theoretical AFLP-PCR experiment and suggest the adaptors and primers needed in the Amplified Fragment Length Polymorphism PCR (AFLP-PCR). A introduction from the site. ... biowww