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Analysis of DNA by Restriction Enzyme Cleavage and Agarose Gel Electrophoresis protocol
A basic protocol of Analysis of DNA by Restriction Enzyme Cleavage and Agarose Gel Electrophoresis for beginners. (David Ng) ...

Optimizing DNA for transformation protocol
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 ...

Restriction Digestion Methods protocol
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 ...

Restriction Enzyme Digestion Troubleshooting Guide new recommended troubleshooting
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 ...

Double restriction digestion of DNA troubleshooting
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 ...

Restriction enzyme digestion troubles troubleshooting
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 ...

Rsa digestion. troubleshooting
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 ...

Labelling of oligos: an interesting phenomenon troubleshooting
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. ... ... ...

Non-radioactive labelling of DNA troubleshooting
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 ...

High temp. restriction enzyme digests troubleshooting
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 ... ... ...

RANDOM primer labelling efficiency troubleshooting
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. ... ... ...

Are my restriction enzymes cutting? troubleshooting
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 ...

Factors that Influence Restriction Enzyme Activity new troubleshooting
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, ...

REBASER: The Restriction Enzyme Database recommended database
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 ...

Integrated Enzyme Database (IntEnz) recommended database
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 ...

RestrictionMapper - with Virtual Digest software
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 ...

NEBCutter 2.0 restriction map and ORF drawing software
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 ...

In silico AFLP-PCR amplification: Suggestion of restriction enzyme pairs new software
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. ...

More restriction enzyme digestion protocols

Figure. Relative positions of different DNA forms of plasmid on agarose gel
(Donis-keller lab manual)
Uncut plasmid DNA can be in any of five forms - nicked circular linear covalently closed supercoiled or circular single-stranded

Protein precipitation after dialysis
(Forum)
dialysed o/n in dialisys buffer contained 50mM Tris pH 8.0, 50 mM NaCl

Secondary Ab background
(Forum)
Secondary antibody caused high background for western blot


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