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小木虫(金币+0.5):给个红包，谢谢回帖
西瓜(金币+5, EPI+1): 学习了！ 2011-07-19 16:25:52
你说的是biological replicate 和 technical replicate，一般每个样本做三到四个技术重复，这个貌似是荧光定量必需的
生物学重复就是可以独立重复提取两三次，分别进行实验，荧光定量最好需要生物学重复～～
另外RNA量最好定在一个范围里面，虽然有内参基因校正，但是大部分综述推荐，这样好调整模版量，并且由于是扩增反应，在2ΔΔct如果扩增效率不平行，很可能误差会放大，因此ΔΔct也最好都做个标准曲线，样品浓度最好在标准曲线范围内。
摘自google～
A technical replicate involves the multiple labeling or reciprocal labeling of the same RNA sample. The purpose of a technical replicate is to control for technical variability within an experiment (array to array variation, reagent variation, dye incorporation, etc.) A well run experiment should have minimal variation thus minimizing the need for technical replicates.
A biological replicate involves isolating RNA independently from replicate sources (multiple cell lines, multiple biopsies, multiple patients, etc). The purpose of a biological replicate is to control for biological diversity. Biological replicates are often more telling, and for this reason are “better” than technical replicates, however, biological replicates are often more difficult to obtain
[ Last edited by shaquila on 2011-7-19 at 13:47 ] |
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