کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
2091167 1081536 2006 8 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Discrimination of viable Vibrio vulnificus cells from dead cells in real-time PCR
موضوعات مرتبط
علوم زیستی و بیوفناوری بیوشیمی، ژنتیک و زیست شناسی مولکولی بیوتکنولوژی یا زیست‌فناوری
پیش نمایش صفحه اول مقاله
Discrimination of viable Vibrio vulnificus cells from dead cells in real-time PCR
چکیده انگلیسی

Ethidium bromide monoazide (EMA) was utilized to selectively allow the real-time PCR (RT-PCR) amplification of a targeted DNA sequence in viable but not dead cells of Vibrio vulnificus. The optimized light exposure time to achieve cross-linking of DNA by the EMA in dead cells and to photolyse the free EMA in solution was at least 15 min. The use of 3.0 μg/ml or less of EMA did not inhibit the PCR amplification of DNA derived from viable cells of V. vulnificus. The minimum amount of EMA to completely inhibit the RT-PCR amplification of DNA derived from heat-killed cells was 2.5 μg/ml. Amplification of DNA from dead cells in a mixture with viable cells was successfully inhibited by 2.5 μg/ml of EMA, whereas the DNA from viable cells present was successfully amplified by RT-PCR.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Journal of Microbiological Methods - Volume 64, Issue 1, January 2006, Pages 1–8
نویسندگان
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