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. 1988 Aug 11;16(15):7351–7367. doi: 10.1093/nar/16.15.7351

A general method of in vitro preparation and specific mutagenesis of DNA fragments: study of protein and DNA interactions.

R Higuchi 1, B Krummel 1, R K Saiki 1
PMCID: PMC338413  PMID: 3045756

Abstract

Specific, end-labeled DNA fragments can be simply and rapidly prepared using the polymerase chain reaction (PCR). Such fragments are suitable for use in DNase I protection footprint assays, chemical sequencing reactions, and for the production and analysis of paused RNA polymerase transcription complexes. Moreover, a general means of introducing a specific mutation at any position along the length of such PCR-generated fragments is described. These procedures, which can circumvent the need for large-scale phage or plasmid growths, preparative gel-electrophoresis and the screening of molecular clones, can facilitate the rapid study of sequence-specific interactions of proteins and DNA. A rapid means of removing excess oligonucleotide primers from completed PCRs is also described.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

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