caa a22 4500 397552378 CHVBK 20180308164753.0 cr unu---uuuuu 161202e199602 xx s 000 0 eng 10.1111/j.1574-6941.1996.tb00205.x doi (NATIONALLICENCE)oxford-10.1111/j.1574-6941.1996.tb00205.x Optimization of conditions for the polymerase chain reaction amplification of DNA from culturable and nonculturable cells of Vibrio vulnificus [Elektronische Daten] [Shirley S. Coleman, James D. Oliver] A series of 16 buffers, differing in pH and MgCl2 concentration, were used to optimize the polymerase chain reaction (PCR) amplification of a 388 bp region of the hemolysin/cytolysin gene from cells of Vibrio vulnificus present in both the culturable and nonculturable states. Both the opaque and translucent morphotypes were examined. Using whole cell lysates, we were able to obtain amplification of DNA from as few as 28.5 cells present in the viable but nonculturable state. With one exception, all buffers that produced amplification using culturable cells also produced amplification using nonculturable cells. However, regardless of the buffer employed, 100 times more nonculturable cells than culturable cells were required to obtain a PCR product. Our data suggest that caution should be exercised when employing PCR optimized against culturable cells when this method is employed for the detection of nonculturable cells. © 1996 Federation of European Microbiological Societies. All rights reserved Polymerase chain reaction nationallicence Vibrio vulnificus nationallicence Viable but nonculturable nationallicence Coleman Shirley S. Department of Biology, University of North Carolina at Charlotte, Charlotte, NC 28223, USA aut Oliver James D. Department of Biology, University of North Carolina at Charlotte, Charlotte, NC 28223, USA aut FEMS Microbiology Ecology Blackwell Publishing Ltd 19/2(1996-02), 127-132 19:2<127 1996 19 femsec https://doi.org/10.1111/j.1574-6941.1996.tb00205.x text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1111/j.1574-6941.1996.tb00205.x text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Coleman Shirley S. Department of Biology, University of North Carolina at Charlotte, Charlotte, NC 28223, USA aut NATIONALLICENCE 700 1- Oliver James D. Department of Biology, University of North Carolina at Charlotte, Charlotte, NC 28223, USA aut NATIONALLICENCE 773 0- FEMS Microbiology Ecology Blackwell Publishing Ltd 19/2(1996-02), 127-132 19:2<127 1996 19 femsec Metadata rights reserved CC BY-NC-4.0 http://creativecommons.org/licenses/by-nc/4.0 nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-oxford