Document Detail


Rapid and reliable dideoxy sequencing of double-stranded DNA.
MedLine Citation:
PMID:  3007296     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
We report a simple and reliable protocol for nucleotide sequencing using the Sanger dideoxy technique on linearized double-stranded DNA molecules with specific oligonucleotide primers. The method is demonstrated for restriction fragments cloned into the plasmid vectors pSP64 and pSP65 using two vector-specific primers, the M13 reverse primer and a new SP6 primer, flanking the multiple cloning site. Template DNA may be prepared by a rapid alkaline lysis procedure. Mild linearization conditions with the appropriate restriction endonuclease avoid the appearance of artifact bands.
Authors:
R G Korneluk; F Quan; R A Gravel
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Gene     Volume:  40     ISSN:  0378-1119     ISO Abbreviation:  Gene     Publication Date:  1985  
Date Detail:
Created Date:  1986-04-25     Completed Date:  1986-04-25     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  7706761     Medline TA:  Gene     Country:  NETHERLANDS    
Other Details:
Languages:  eng     Pagination:  317-23     Citation Subset:  IM    
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MeSH Terms
Descriptor/Qualifier:
Base Sequence
Cloning, Molecular
DNA Restriction Enzymes
DNA, Recombinant*
Genetic Vectors
Phosphorus Radioisotopes
Plasmids
Templates, Genetic
Chemical
Reg. No./Substance:
0/DNA, Recombinant; 0/Phosphorus Radioisotopes; EC 3.1.21.-/DNA Restriction Enzymes

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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