One-tube restriction enzyme digest and fluorescent labeling for restriction endonuclease fingerprinting single-strand conformational polymorphism

One-tube restriction enzyme digest and fluorescent labeling for restriction endonuclease fingerprinting single-strand conformational polymorphism.

Fluorescent substrates for the EcoRV restriction endonuclease.

In the presence of Mg2+, the EcoRV restriction endonuclease cleaves DNA specifically at the sequence GAT'ATC (where denotes the point of scission) [ I ] . Even the change of a single base pair within this sequence will lead to a million fold reduction of EcoRV activity [2]. Paradoxically, gel retardation experiments in the absence of Mg2+ show that, unlike EcoRl and many other type 11 restricti...

Intraspecies polymorphism of Cryptosporidium parvum revealed by PCR-restriction fragment length polymorphism (RFLP) and RFLP-single-strand conformational polymorphism analyses.

A glycoprotein (Cpgp40/15)-encoding gene of Cryptosporidium parvum was analyzed to reveal intraspecies polymorphism within C. parvum isolates. Forty-one isolates were collected from different geographical origins (Japan, Italy, and Nepal) and hosts (humans, calves, and a goat). These isolates were characterized by means of DNA sequencing, PCR-restriction fragment length polymorphism (PCR-RFLP),...

REF Select: expert system software for selecting restriction endonucleases for restriction endonuclease fingerprinting.

REF Select, expert system software, has been developed to assist in the selection of optimal restriction endonucleases for restriction endonuclease fingerprinting (REF), a method for rapid and sensitive mutation screening of long DNA segments (1-2 kb). The REF method typically involves six separate digestions with up to two restriction endnonucleases used in each digestion. If done manually, pe...

Dual restriction enzyme digest of cationic-gold-coated DNA scaffolds

DNA strands coated with AuNPs were cleaved by restriction enzymes while in solution or on a surface. Enzymatic activity was verified by gel electrophoresis prior to surface analysis. Cleavage results suggest that enzymes can recognize the AuNP-coated strands while on the surfaces, though specificity in digestion has not yet been verified. Development allows for advances in site specific localiz...