赛默飞世尔Thermo Fisher X125 8X0,2M

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

Counting ES cell Chromosomes

for four hours in the presence of colcemid (3.125 µl of 10 µg/ml colcemid (Gibco # 15212-012) per ml of medium). 2ml per 35 mm and 4ml per 60 mm. 3) Remove media and place it in a 15 ml Falcon tube. Wash with trypsin, and trypsinize with 1.5 ml trypsin.

RNA purification using TRizol--TRIZOL法提取RNA

of chloroform to each tube.b.Cap each tube. Shake samples vigorously by hand for 15 seconds.c.Incubate samples for 5 minutes at room temperature.d.Centrifuge samples for 15 minutes at 12,000 x g at 4oC.3.  RNA Precipitation a.Transfer the upper aqueous phase