"Nonrandom" DNA sequence analysis in bacteriophage M13 by the dideoxy chain-termination method.
Open Access
- 1 July 1982
- journal article
- research article
- Published by Proceedings of the National Academy of Sciences in Proceedings of the National Academy of Sciences
- Vol. 79 (14), 4298-4302
- https://doi.org/10.1073/pnas.79.14.4298
Abstract
We describe a rapid "nonrandom" DNA sequence analysis procedure that facilitates the nucleotide sequence determination of large contiguous regions of DNA. The method consists of cloning a restriction endonuclease fragment of interest into bacteriophage M13 followed by construction of a series of nuclease BAL-31 deletion mutants originating from a single site in M13 that is close to the DNA insert. Determination of the size of the deletion mutant is accomplished by hybridization to a complementary single-stranded probe derived from M13 containing that total insert followed by nuclease S1 treatment. Single-stranded M13-insert DNAs of progressively smaller sizes are isolated and analyzed by using a site-specific M13 DNA primer and the dideoxy chain-termination method. In this way, analysis of the DNA sequence proceeds from one end of the total insert to the other in a nonrandom fashion due to generation of a controlled overlapping set of deletion mutants.This publication has 17 references indexed in Scilit:
- A versatile primer for DNA sequencing in the M13mp2 cloning systemGene, 1980
- Isolation, characterization, and comparison of recombinant DNAs derived from genomes of human hepatitis B virus and woodchuck hepatitis virus.Proceedings of the National Academy of Sciences, 1980
- Repetitive DNA sequences near three humanβ-type globin genesNucleic Acids Research, 1980
- A rapid alkaline extraction procedure for screening recombinant plasmid DNANucleic Acids Research, 1979
- DNA sequence from the histidine operon control region: seven histidine codons in a row.Proceedings of the National Academy of Sciences, 1978
- The use of thin acrylamide gels for DNA sequencingFEBS Letters, 1978
- DNA sequencing with chain-terminating inhibitorsProceedings of the National Academy of Sciences, 1977
- Filamentous coliphage M13 as a cloning vehicle: insertion of a HindII fragment of the lac regulatory region in M13 replicative form in vitro.Proceedings of the National Academy of Sciences, 1977
- A new method for sequencing DNA.Proceedings of the National Academy of Sciences, 1977
- An electron microscope study of the DNA sequence organization of the human genomeJournal of Molecular Biology, 1976