Publication Date:
1992-12-11
Description:
When template DNA is saturated with a single-stranded DNA binding protein (SSB), strings of three or four contiguous hexanucleotides (hexamers) can cooperate through base-stacking interactions to prime DNA synthesis specifically from the 3' end of the string. Under the same conditions, priming by individual hexamers is suppressed. Strings of three of four hexamers representing more than 200 of the 4096 possible hexamers primed easily readable sequence ladders at more than 75 different sites in single-stranded or denatured double-stranded templates 6.4 kilobases to 40 kilobase pairs long, with a success rate of 60 to 90 percent. A synthesis of 1 micromole of hexamer supplies enough material for thousands of primings, so multiple libraries of all 4096 hexamers could be distributed at a reasonable cost. Such libraries would allow rapid and economical sequencing. Automating this strategy could increase the speed and efficiency of large-scale DNA sequencing by at least an order of magnitude.〈br /〉〈span class="detail_caption"〉Notes: 〈/span〉Kieleczawa, J -- Dunn, J J -- Studier, F W -- New York, N.Y. -- Science. 1992 Dec 11;258(5089):1787-91.〈br /〉〈span class="detail_caption"〉Author address: 〈/span〉Biology Department, Brookhaven National Laboratory, Upton, NY 11973.〈br /〉〈span class="detail_caption"〉Record origin:〈/span〉 〈a href="http://www.ncbi.nlm.nih.gov/pubmed/1465615" target="_blank"〉PubMed〈/a〉
Keywords:
*Base Sequence
;
Binding Sites
;
DNA/*genetics/metabolism
;
DNA, Viral/genetics
;
DNA-Binding Proteins/metabolism
;
Escherichia coli/metabolism
;
*Genetic Techniques
;
Molecular Sequence Data
;
Nucleic Acid Denaturation
;
Oligodeoxyribonucleotides
;
Sulfur Radioisotopes
;
Templates, Genetic
Print ISSN:
0036-8075
Electronic ISSN:
1095-9203
Topics:
Biology
,
Chemistry and Pharmacology
,
Computer Science
,
Medicine
,
Natural Sciences in General
,
Physics
Permalink