伯乐基因表达分析方法学部分

1、Gene Expression Analysis for iCycler iQ Real-Time PCR Detection System produce a final composite standard deviation for the expression level according to accepted procedures for error propagation. For details, see Vandesompele et al., Genome Biology 2002 and the geNorm website: http:/medgen.ugent.be

2、/jvdesomp/genorm/step ensures that final expression for the control. The scaled expressdividing the unscaled level by the ucontrol sample. If no control is spescaled relative to the lowest unscasame gene as the sample of intereIn the example, the unscaled expreid cat01, is unchanged by scalingexpres

3、sion level of 1.0 for the contcontrol-id.Detection System2004, Bio-Radxpression in this spreadsheet are:he same gene, type and identifier, sample. This mean and its standard output data grid, shown here withere, the Ct values for the two wells, (i.e. blank type and identifier cat01, to produce a mea

4、n Ct value of 26.70.e a quantity relative to the control no control is specified the quantity is ple for the same gene with the lowest lative quantity is:(control C t -unknown Ct cyfor gene A, id cat01, has been or the matching control sample for duce a quantity for gene A, id cat01,t, calculate a n

5、ormalization factor, of the relative quantities for all ame identifier as that sample. Theant. 1 * quant. 2 * . . .* quant. NN(1samples for gene A, id cat01, are r genes b, c and d. The relative es are combined using the geometric zation factor, which is not displayed.ression level for the sample of

6、 interestexpression level of 0.074 for gene A, iding its relative quantity by the rom the previous step.sion level for the sample of interest trol sample for the same gene. This sion levels are relative to a value of 1 sion levels are relative to a value of 1 xpression level is calculated bythe unscaled expression level of the s specified, the expression level is unscaled expression level for the nterest.expr