Some modifications in the procedure of direct sequencing of PCR amplified 16S rDNA

Dorsch M. and Stackebrandt E. (1992) Some modifications in the procedure of direct sequencing of PCR amplified 16S rDNA. Journal of Microbiological Methods, 16 4: 271-279. doi:10.1016/0167-7012(92)90017-X


Author Dorsch M.
Stackebrandt E.
Title Some modifications in the procedure of direct sequencing of PCR amplified 16S rDNA
Journal name Journal of Microbiological Methods   Check publisher's open access policy
ISSN 0167-7012
Publication date 1992-01-01
Sub-type Article (original research)
DOI 10.1016/0167-7012(92)90017-X
Volume 16
Issue 4
Start page 271
End page 279
Total pages 9
Subject 1305 Biotechnology
2402 Applied Microbiology and Biotechnology
2404 Microbiology
Abstract A method for the analysis of PCR-amplified 16S ribosomal (r)DNA has been developed that generates sequences of excellent quality. In order to avoid time consuming purification and cloning steps and to reduce the number of non-specific terminations that frequently occur during direct sequencing of 16S rDNA, we developed a strategy that combines the advantages of several procedures described previously for different purposes, such as purification of amplified DNA via the Prep-A-Gene method and the snap-freeze technique in the sequencing protocol. Using 40-400 ng DNA per PCR reaction the amount of PCR product was sufficient to subsequently generate a complete 16S rDNA sequence. The primary structures of these genes were determined fast and reliably, yielding sequences in which the rate of unidentified positions usually did not exceed 0.2%.
Keyword 16S rRNA gene amplification
DNA sequencing
Phylogenetic studies
Q-Index Code C1
Q-Index Status Provisional Code
Institutional Status Unknown

Document type: Journal Article
Sub-type: Article (original research)
Collection: Scopus Import
 
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Created: Sat, 09 Jul 2016, 17:38:29 EST by System User