Please use this identifier to cite or link to this item: https://doi.org/10.1002/bit.10552
Title: Flow cytometric detection of β-D-glucuronidase gene in wild-type bacterial cells using in-situ PCR
Authors: Sachidanandham, R. 
Gin, K.Y.-H.
Keywords: Coliform
E. coli
Flow cytometry
ISPCR
Uid
Issue Date: 20-Apr-2003
Citation: Sachidanandham, R., Gin, K.Y.-H. (2003-04-20). Flow cytometric detection of β-D-glucuronidase gene in wild-type bacterial cells using in-situ PCR. Biotechnology and Bioengineering 82 (2) : 127-133. ScholarBank@NUS Repository. https://doi.org/10.1002/bit.10552
Abstract: An in situ PCR-based flow cytometry method useful for monitoring the presence or absence of the β-D-glucuronidase gene in Escherichia coli has been developed. A single-step fixation and permeabilization procedure, which maintained cell integrity at the elevated temperatures used during thermal cycling in the presence of PCR reagents, was demonstrated. We have chosen a shorter DNA sequence of length 147 bp for the PCR. Cells subjected to in situ PCR using fluorescein-12-dUTP as a label, showed the presence of uid both in epifluorescence microscopic examination and flow cytometric analysis. Multi-parametric analysis of flow cytometric profiles revealed that the efficiency of labeling was found to be high. The potential of in situ PCR for the detection of uid in intact coliform cells was then successfully tested with a fecal coliform isolated from the coastal waters of Singapore. © 2003 Wiley Periodicals, Inc.
Source Title: Biotechnology and Bioengineering
URI: http://scholarbank.nus.edu.sg/handle/10635/132785
ISSN: 00063592
DOI: 10.1002/bit.10552
Appears in Collections:Staff Publications

Show full item record
Files in This Item:
There are no files associated with this item.

SCOPUSTM   
Citations

8
checked on Nov 14, 2018

WEB OF SCIENCETM
Citations

7
checked on Nov 14, 2018

Page view(s)

20
checked on Nov 1, 2018

Google ScholarTM

Check

Altmetric


Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.