Comparison of RT-qPCR approaches for quantification of cyanobacteria in Lake Taihu
To evaluate the real-time quant itative PCR quanit ification(RT-qPCR) approaches for determining cyanobacteria and Microcystis,four methods of SYBR Green I RT-qPCR were established based on the target genes,including the 16S rRNA gene of cyanobacteria,the phycocyanin and the 16S rRNA gene of Microcystis.Two different standards including M.aeruginosa PCC 7806 genomic DNA and plasmid DNA carrying the phycocyanin gene were utilized.Cultured Microcystis aeruginosa and environmental samples of Lake Taihu were applied to evaluate the four methods from which the results were compared with that of microscope counting.In general, there was good agreement between the cell number equivalents obtained from RT-qPCR assay and microscope counting for the cultured Microcystis aeruginosa test, but for environmental samples only the results of RT-qPCR assay using M.aeruginosa PCC 7806 genomic DNA as standard and phycocyanin gene as the target gene were consistent with those estimated via the microscope.So as a monitoring method, RT-qPCR analysis could meet the requirement of algae bloom quantification and it was convenient,rapid and specific.
PENG Yu-Ke YUE Dong-Mei WU Jun XIAO Lin YANG Liu-Yan
State Key Laboratory of Pollution Control and Resource Reuse ,School of the Environment ,Nanjing University ,Nanjing ,Jiangsu 210046,China
国际会议
武汉
英文
14
2011-04-05（万方平台首次上网日期，不代表论文的发表时间）