作者
Himanshu Mali, Chandni Shah, Anil S Prajapati, Sureshkumar Mesara, Hiren A Dhameliya, Darshan H Patel, Ujjval Trivedi, RB Subramaniam
发表日期
2022/2
期刊
Applied Microbiology and Biotechnology
卷号
106
期号
4
页码范围
1705-1714
出版商
Springer Berlin Heidelberg
简介
Organophosphates are becoming an emerging pollutant due to their various applications, particularly as pesticides. In this study, an improved Colony (Live-cell) PCR method was developed for the detection of opd genes from bacteria encoding the organophosphate hydrolase enzymes capable of degrading various organophosphates. The improved method does not require pre-heating or pre-lysis of bacterial cells as essential in the conventional colony PCR. The reaction volume was scaled down to 10 µl by optimizing the PCR buffer and amplification conditions. The improved method was used for Gram positive and negative bacteria, glycerol stocks, liquid cultures, recombinant and mutant strains. Also, 16S rRNA gene was amplified from unknown environmental isolates and known E. coli strains. The amplified opd and 16S rRNA genes from the improved colony PCR method and by conventional PCR were …
学术搜索中的文章
Improved live-cell PCR method for detection of organophosphates degrading opd genes and applications
H Mali, C Shah, AS Prajapati, S Mesara, HA Dhameliya… - Applied Microbiology and Biotechnology, 2022