The sensitivity of the real-time PCR procedure was evaluated at 500 HCMV DNA copies per ml of sera. The use of an internal control concomitantly processed during the HCMV quantification did not alter the sensitivity of the procedure, and was relevant for the detection of putative PCR ...
Development of a real time-PCR procedure for the quantification of cumulus cell mrnas before oocyte inseminationOBJECTIVE: To develop a procedure for quantitative analysis of gene expression in cumulus cells (CC) that could be completed within 3 hours to help embryologist selecting the best oocytes...
Step 3: Detection Using Real-time PCR. Finally, 10 µL of each reverse transcription reaction served as template in a real-time PCR procedure using one of 16 TaqMan Gene Expression Assays (Figure 4). Each 100 µL lysate ...
In this chapter, an in-house HIV-1 real-time RT-PCR assay for blood donor screening is described. The procedure includes the pooling of plasma samples, viral RNA isolation, and subsequent detection of amplification in real-time one-step RT-PCR. The validation considers the specificity, the ...
time quantitative PCR procedure in this outbreak incidence,and then 5 isolates of Sh.flexneri were isolated from the enrichment broth.The PFGE patterns of the 5 strains were identical,which expressed the high homology of them.[Conclusion]Real-time PCR procedure is a method with high sensitivity,...
PCR-based methods could be a good alternative to traditional culturing techniques since they are rapid, sensitive, specific and allow accurate quantification. The aim of this study was to optimize the conditions for a real-time PCR method to enumerateL. monocytogenesin cheese, and the applicability...
由于后续只针对Real-timePCR阳性标本进行检测,减少了实验者的工作量,节约了阴性标本后续检测的实验试剂.%To improve the detection rate of Salmonella by using Real-time PCR, and to simplify the experimental procedure and reduce the amount of work, so as to provide the experimental evidence for the ...
Real-time RT-PCR has become a common technique, no longer limited to specialist core facilities. It is in many cases the only method for measuring mRNA levels of vivo low copy number targets of interest for which alternative assays either do not exist or lack the required sensitivity. Benefi...
[44]. Therunning process of the real-time PCR can be monitored directly by the exponential curve analysis on a computer screen[45]. The procedure initially require reverse transcriptase enzyme to convert the single strand RNA to cDNA through primary PCR[46]. Experimental approach to this ...
real-time PCR将会影响 PCR技术的发展,并引起生物学研究者,尤其是分子诊断学、 分子生理学和科学教育研究者的极大兴趣。 1real-time PCR的原理 其原理是利用能特异标记PCR产物的荧光物质,利用荧光信号积累实时监测整个PCR进程,得到S型的扩增曲线。假设该曲线的前期PCR符合指数性扩增,于是在单纯指数方程的基础上,通过...