切片石蜡50-52°c

RNA-biotin based pulldown assays for the detection of siRNA targeted genomic regions and siRNA directed histone modifications (PROT32)

so that there are no clumps of cells. Incubate on ice for 10 minutes;Centrifuge at 5,000 rpm for 5 minutes at 4°C to pellet the nuclei;Re-suspend nuclei in 600µl of ChIP lysis buffer+1/1000 PMSF+50 units of RNAse inhibitor and incubate on ice for 10 minutes; Sonicate

纯化后的蛋白质怎样保存？

0.22 µm 过滤器），或者加入抑菌剂（例如 0.1% 叠氮化钠）以避免细菌生长。值得注意的是，并非所有蛋白质都能在 4°C 长时间保持稳定。 3. 长期保存（一周以上） 需要将蛋白样品冻存。使用液氮或者干冰/乙醇混合物将其快速冷冻避免蛋白质变性。可以将样品分装至多个管子中，来避免反复冻融可能造成的活性降低或影响结构。 可以添加几种稳定剂，如甘油（5 - 50%（w / v）），血清白蛋白（10 mg / ml），还原剂（例如 1 mM DTT）和配体（其性质和浓度取决于靶蛋白的性质和浓度

Primer Premier: Program for Design of Degenerate Primers from a Protein Sequence

mixture was denatured for 7 min at 95°C and subjected to 28 cycles each with a profile of 60 s at 92°C, 80 s at 52°C and 105 s at 72°C in a DNA Thermal Cycler (Perkin-Elmer, Norwalk, CT, USA). The electrophoresis of the PCR product was performed on a 1