全基因组测序以及转录组数据库的建立为系统地研究基因功能提供可能。目标基因表达数据为了解其生物学功能提供了线索。实时定量PCR(RT-qPCR)是快速研究基因功能的良好方法。这个方法需要稳定表达的参考基因(reference gene)来均一化数据从而得到精确可靠的数据结果。
Reference genesGeNormTriticum aestivumPuccinia graminis f.sp triticiPuccinia triticinaPuccinia striiformisReal-time PCR (qPCR) is an effective method to quantify mRNA levels, but requires validated reference genes for data normalisation. The GeNorm-Plus algorithm was used to examine the expression stability...
The stability of the reference genes showed different results according to the chemicals (Fig.1, Fig.S2-S5). Based on the M values evaluated by geNorm, all genes and chemicals showed M values of less than 1, an acceptable value considering suitable reference genes for RT-qPCR normalization in...
To validate the selected reference genes for qRT-PCR normalization, the relative expression profiles of theMEP3gene were analyzed.MEP3is known as the gene encodingM. caniskeratynolityc metalloprotease (43.5 kDa) expressed in the presence of keratin26. The validation analysis was conducted using ...
Validation of reference genes for quantitative RT-qPCR studies of gene expression in Atlantic cod ( Gadus morhua l .) during temperature stress. Ingrid A Aursnes,Anne L Rishovd,Hans E Karlsen,Tor Gj?en. BMC Research Notes . 2011Aursnes, I. A., Rishovd, A. L., Karlsen, H. E. & ...
(RT—qPCR)isatechniquethatiscurrentlyusedextensively.Reliablequantificationdependsontheselectionofoneormorestablyexpressedreferencegene(s)(RGs),whichareusuallyhousekeepinggenes,asinternalcon-trols.Theyincludegenesencodingproductswithfunctionsthatmaintainthecell—wallstructureandtheprimarymetabolism,suchas18SribosomalRNA(...
To ensure comparability between the 14 (13 reference genes + interleukin-2) QPCR assays, we first determined the PCR efficiency of each individual assay by measuring serial dilutions of 100 ng cDNA from CCRF-HSB-2 cells in triplicate [20]. Inter-assay variation was investigated in three ...
To lessen bias induced by an unsuitable reference gene, many researchers have recommended the use of two or more reference genes for qPCR study [3, 33–35]. For the developmental study in seven tissues, we chose two sets of genes, e.g., one set of HPRT1 and 18S (the two most ...
(RT-qPCR)hasmainlybeenwidelyappliedtodetectgeneexpressionbecauseo itssimplicity, astness,lowcostandhighsensitivity.Oneo therequirements orRT-qPCRistheavailabilityo suitablere erencegenes ornormalizationo geneexpression.However,currently,noPassi oraedulisre erencegeneshavebeenidenti edandthusithashinderedthegene...
Identification of appropriate reference genes for qRT-PCR analysis of heat- stressed mammary epithelial cells in riverine buffaloes (Bubalus bubalis). ISRN ... N Kapila,A Kishore,M Sodhi,... 被引量: 13发表: 2013年 MiR-15a decreases bovine mammary epithelial cell viability and lactation and reg...