Abstract：
:A real-time PCR (RTm-PCR) assay using fluorescently labeled oligonucleotides (TaqMan probes) was used to detect and quantify the recombinant Rhodococcus sp. strain RHA1(fcb) in soil. One primer and probe set targeted a hypervariable region of the 16S rRNA gene unique to strain RHA1(fcb) and its phylogenetic relatives, and the other set targeted the recombinant 4-chlorobenzoate (4-CBA) degradation operon (fcb) and was strain-specific. The method had a 6-log dynamic range of detection (10(2)-10(7) cells ml(-1)) for both probes when DNA from pure cultures was used. Although the method was less sensitive in soil, the estimated number of cells in soil by real-time PCR corresponded to the measured number of RHA1(fcb) cells determined by colony-forming units.
journal_name
J Microbiol Methodsjournal_title
Journal of microbiological methodsauthors
Rodrigues JL,Aiello MR,Urbance JW,Tsoi TV,Tiedje JMdoi
10.1016/s0167-7012(02)00067-2subject
Has Abstractpub_date
2002-10-01 00:00:00pages
181-9issue
2eissn
0167-7012issn
1872-8359pii
S0167701202000672journal_volume
51pub_type
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