Triplex affinity capture of a single copy clone from a yeast genomic library
نویسندگان
چکیده
منابع مشابه
Rapid screening of a human genomic library in yeast artificial chromosomes for single-copy sequences.
A yeast artificial chromosome (YAC) library in Saccharomyces cerevisiae consisting of 30,000 clones with an average insert size of 0.1 megabase pair of human DNA has been generated from primary fibroblast DNA. A YAC vector was modified to enable the recovery of both ends of a human DNA insert in plasmids in Escherichia coli and to confer G418 resistance to mammalian cells. A rapid method for ye...
متن کاملAn anonymous single copy clone (p1,2-2) from a flow sorted human chromosomes 1 and 2 library (D1S33).
SOURCE AND DESCRIPTION OF CLONE: pi,2-2, a 1.8 Kb EcoRI fragment was subcloned in pBR322, derived from a genomic library in Charon 21A which was constructed from sorted by flow cytometry human chromosome 1 and 2. POLYMORPHISM: EcoRI indentifies two alleles with DNA fragments at either 1.8 Kb (Al) or 1.0 and 0.3 Kb (A2), Hind III identifies two alleles with DMA fragments at either 5.0 (Bl) or 8....
متن کاملSequence-specific DNA purification by triplex affinity capture.
A DNA isolation procedure was developed by using triple-helix formation and magnetic separation. In this procedure, target DNA is captured by a biotinylated oligonucleotide via intermolecular triplex formation, bound to streptavidin-coated magnetic beads, and recovered in double-stranded form by elution with a mild alkaline buffer that destabilizes the triple helix. The effectiveness of the pro...
متن کاملOptimized Affinity Capture of Yeast Protein Complexes.
Here, we describe an affinity isolation protocol. It uses cryomilled yeast cell powder for producing cell extracts and antibody-conjugated paramagnetic beads for affinity capture. Guidelines for determining the optimal extraction solvent composition are provided. Captured proteins are eluted in a denaturing solvent (sodium dodecyl sulfate polyacrylamide gel electrophoresis sample buffer) for ge...
متن کاملA Chlamydomonas genomic library in yeast artificial chromosomes.
We have constructed and characterized a Chlamydomonas reinhardtii total genomic library in yeast artificial chromosomes (YACs). The library contains 7500 clones with inserts ranging in size from 100-200 kb. The representation of the library was assessed by screening one-third of it with a probe derived from the dispersed repeat, Gulliver, which occurs approximately 13 times in the genome. At le...
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ژورنال
عنوان ژورنال: Nucleic Acids Research
سال: 1992
ISSN: 0305-1048,1362-4962
DOI: 10.1093/nar/20.13.3524