赛默飞世尔Thermo Fisher X25 BAGS,PO

ChIP protocol for X. laevis Lens1/FoxE3 enhancer

(9)  Sonicate the homogenate on ice (10 sec. pulse + 50 sec. break x 6 times, 20% amplitude with a small tip of Branson sonifier (Fisher Dismembranator Model 500)). This step releases chromatin DNA from cells without solubilizing yolk proteins that make

Assessment of Magnaporthe grisea mating type by spore PCR

) containing 50 mM KCl, 0.1 Triton X-100, 2.5 mM MgCl2, 0.2 mM each dNTP, 1.0 mM each primer, 1.25 units of Taq DNA polymerase (Promega). Samples are overlaid with 50 ul mineral oil and subjected to the following PCR condition: 95 C for 5 min; 30 cycles of 95 C

一篇搞定 IP、co-IP 实验

糖，建议进行 Pre-clear；如果使用磁珠，此步可忽略 抗 X 抗体的轻链（25 kD）和重链（50 kD） 更侧重于如何在实验上进行优化（见下文） *注意：总蛋白上样量过多也会造成 非特异性结合，建议上样量为 500 ug-1 mg。上样之前一定要先用 BCA 定个量！定个量！定个量！ pre-clear：上样前先用裂解液过一遍柱子，减少基质本身对蛋白的吸附 Output：在某些 co-IP 实验中，实验人员会把IP后的上清分别进行诱饵蛋白 X 和靶蛋白 Y 的 WB 检测