A short primer for sequencing DNA cloned in the single-stranded phage vector M13mp2 | Zendy

Stephen K. Anderson | Zendy; Michael J. Gait | Zendy; Luciano Mayol | Zendy; Ian G. Young | Zendy

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A short primer for sequencing DNA cloned in the single-stranded phage vector M13mp2

Author(s) -

Stephen K. Anderson,

Michael J. Gait,

Luciano Mayol,

Ian G. Young

Publication year - 1980

Publication title -

nucleic acids research

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 9.008

H-Index - 537

eISSN - 1362-4954

pISSN - 0305-1048

DOI - 10.1093/nar/8.8.1731

Subject(s) - ecori , biology , bamhi , plasmid , microbiology and biotechnology , primer (cosmetics) , cloning vector , restriction site , in vitro recombination , molecular cloning , genetics , multiple cloning site , dna , pbr322 , vector (molecular biology) , restriction enzyme , gene , recombinant dna , peptide sequence , chemistry , organic chemistry

In this paper we describe the synthesis and cloning of a short segment of DNA complementary to the region immediately adjacent to the EcoRI insertion site in the single-stranded bacteriophage vector M13mp2. This segment is useful as a "universal" primer for DNA sequencing by the dideoxynucleotide chain termination method; the template can be any DNA species cloned in M13mp2 or its derivatives. The primer has been cloned into the tetracycline resistance gene of plasmid pBR322 as one strand of a 26 bp EcoRI/BamHI fragment. This fragment may be readily prepared from an EcoRI + BamHI restriction digest of the parent plasmid (designated pSP14) by a simple size fractionation.

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