Amplification of 4–9-kb Human Genomic DNA Flanking a Known Site Using a Panhandle PCR Variant | Zendy

Douglas H. Jones | Zendy; Stanley C. Winistorfer | Zendy

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Amplification of 4–9-kb Human Genomic DNA Flanking a Known Site Using a Panhandle PCR Variant

Author(s) -

Douglas H. Jones,

Stanley C. Winistorfer

Publication year - 1997

Publication title -

biotechniques

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 0.617

H-Index - 131

eISSN - 1940-9818

pISSN - 0736-6205

DOI - 10.2144/97231rr01

Subject(s) - primer dimer , primer extension , oligonucleotide , primer (cosmetics) , hot start pcr , genomic dna , microbiology and biotechnology , polymerase chain reaction , dna , biology , multiple displacement amplification , inverse polymerase chain reaction , genetics , multiplex ligation dependent probe amplification , multiplex polymerase chain reaction , gene , chemistry , base sequence , dna extraction , exon , organic chemistry

We present a method for the in vitro amplification of > 6.0 kb of DNA flanking a known site. This is accomplished by ligating an oligonucleotide to create an inverted repeat of a portion of the known sequence, followed by single-primer polymerase chain reaction (PCR) amplifications. This method generates a panhandle template following primer extension on the strand of interest. It does not involve template-directed extension from the ligated oligonucleotide, and it is carried out without DNA extractions. We have used this method to amplify 4.5-9.4 kb of DNA flanking the original primer annealing sites directly from human genomic DNA.

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