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文献和实验Reducing Sample Complexity by RP-HPLC: Beyond the Tip of the Protein Expression Iceberg
Because the dynamic range of most cell or tissue proteomes is enormous, separation of such complex protein samples by two-dimensional electrophoresis (2-DE) on broad pH gradients often results in the visualization of only the most abundantly
Clonality - X Chromosome Inactivation Assay
for 5 min at 94°C. Load 3 µl onto gel. Electrophorese at 1800 volts for 1 to 2 hours. TIP: In clonality analysis, the PCR products are ~200 bp. Therefore do not overload the gel with sample and run the gel for a longer period of time to sharpen
做 ChIP(Chromatin Immunoprecipitation)已经几个月了,虽时间不长,但深刻体会细节决定成败,这里就把自己认为值得注意的细节总结如下: 1、cell counting:尽量做到准确,会影响 input 结果。 2、cross link:甲醛的终浓度是 1%,这个基本所有的 protocol 上都会强调。 3、resuspend cells with SDS:一定要选用小的 tip 头,在液面下吹打,否则很容易产生气泡,后面的 sonication 就麻烦
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