Open AccessA short primer for sequencing DNA cloned in the single-stranded phage vector M13mp2
Author(s) -
Stephen 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.