Dwivedi P.P., Patel B.K.C., Rees G.N., Ollivier Bernard. (1996). A rapid method for sequencing of rRNA gene(s) amplified by polymerase chain reaction using an automated DNA sequencer. Indian Journal of Microbiology, 36, p. 9-12.
Titre du document
A rapid method for sequencing of rRNA gene(s) amplified by polymerase chain reaction using an automated DNA sequencer
Année de publication
1996
Type de document
Article
Auteurs
Dwivedi P.P., Patel B.K.C., Rees G.N., Ollivier Bernard
Source
Indian Journal of Microbiology, 1996,
36, p. 9-12
A method for DNA sequencing of ribosomal RNA (rRNA) genes, amplified by polymerase chain reaction (PCR), using internal primers, designed on the basis of conserved regions of rRNA genes for determining a near complete sequence (99%) of the gene using an automated DNA sequencer (Applied Biosystem Incorporation, USA) is described. The procedure is extremely rapid as cloning of the gene is not required for sequence determination. In addition time consuming steps such as ethanol precipitation and hazardous steps such as phenol/chloroform extractions are excluded from the protocol for the purification of extension products after Taq cycle sequencing using the ABI dye terminator chemistry. The method has been successfully used for sequencing of the 16S and 18S rRNA genes of microbes which includes six members of domain Bacteria, one of domain Archaea and one belonging to Eukarya domain. (Résumé d'auteur)
Plan de classement
Biologie [020BIOL]
Descripteurs
BIOLOGIE MOLECULAIRE ; GENE ; ARN ; METHODE D'ANALYSE ; PCR.REACTION DE POLYMERISATION EN CHAINE
