PCR-INTRODUCED LOOP STRUCTURE AS PRIMER IN DNA-SEQUENCING

Citation
M. Ronaghi et al., PCR-INTRODUCED LOOP STRUCTURE AS PRIMER IN DNA-SEQUENCING, BioTechniques, 25(5), 1998, pp. 876
Citations number
17
Language
INGLESE
art.tipo
Article
Categorie Soggetti
Biology,"Biochemical Research Methods
Journal title
BioTechniques → ACNP
ISSN journal
0736-6205
Volume
25
Issue
5
Year of publication
1998
Database
ISI
SICI code
0736-6205(1998)25:5<876:PLSAPI>2.0.ZU;2-V
Abstract
The need for a primer hybridization step before sequencing has been el iminated using a stem-loop structure generated by PCR. The loop struct ure is obtained by careful design of the PCR primer or by cloning the target DNA into a dedicated vector (pRIT 28HP). After solid-phase capt ure of the PCR product, the loop is formed by elution of the non-bound strand. Here, Mit shaw that both the immobilized ann the eluted stran d call be analyzed using conventional Sanger DNA sequencing and the no vel pyrosequencing method as described previously. By using a stem-loo p structure as a primer for DNA sequencing, the risk fbr mispriming is minimized.