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

25 April 1980

journal article
research article
Published by Oxford University Press (OUP) in Nucleic Acids Research

Vol. 8 (8) , 1731-1743
https://doi.org/10.1093/nar/8.8.1731

Abstract

In this paper we describe the synthesis and cloning of a short segment of DNA complementary to the region immediately adjacent to the Eco RI 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 Eco RI/ Bam HI fragment. This fragment may be readily prepared from an Eco RI + Bam HI restriction digest of the parent plasmid (designated pSPi4) by a simple size fractionation.

Keywords

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