Different Experimental Method Detection GelE Sensitivity Factor Differences

Purpose: Through the use of common two-step RT-PCR and Real-time PCRdetection Enterococcus faecalis of gelE factor expression, evaluation Real-time PCRtechnology determination dung bowel coccus mRNA expression level of the value.Methods:Eextraction Enterococcus faecalis standard strains total RNA genome,the total RNA genome DNA enzymes processing, determination of total RNA genomeconcentration, through the retrovirus PCR will RNA retrovirus get cDNA, gradientdegree evaluation Real-time PCR technology determination dung bowel coccus mRNAexpression level of the value.Results: RT-PCR detection is with the biggest positive amplification channel "fordilution degree3, gene expression sensitivity for550pg/ul. Real-time PCR in dilute1:10for5is still present positive amplification curve, the gene expression sensitivity of5.5pg/ul, Real-time PCR on determination of dung bowel coccus poisonous force factorgelE sensitivity is lower conventional RT-PCR100times.Conclusion:1RT-PCR and Real-time PCR detection can be as dung bowelcoccus mRNA the quantitative analysis method.2detection dung bowel coccus poisonous force of gelE factor method, Real-timePCR method, the sensitivity of relatively common two-step RT-PCR high100times.