2'-Methyl-adenosine-(Bz)-succi

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

upon the dual pulldown to incorporate a third pulldown which is an iteration of the ChIP and is a pulldown for H3K27me3+ (Figure 1b). The third assay described here is the biotin-RNA pulldown of a low-copy RNA that spans the siRNA targeted promoter region

96-well RNA In Situ Hybridization Protocol

. to overnight). To stop development of individual wells, remove staining solution and add 200μlPBT. 26. To stop entire plate, rinse plate 3X with PBT. 27. Add 200μl70% gycerol to each well. 28. Embryos are ready to mount or dissect once they have settled

RNA的电泳，转膜和杂交

1hr左右，至溴酚兰离点样孔约3.5-4cm（凝胶在紫外灯下可看到28S和18S刚好分开即可） 6.转膜：转膜液用500ml 4×SSC加27ml 37%甲醛（终浓度为2%）；转膜步骤同Southern。注意加溶液后一切操作均在通风橱内进行，以避免甲醛对人体的伤害 7.照相：将胶及膜取下，分别在凝胶成像系统下看胶上RNA是否有残留，膜上RNA效果是否完好。 8.风干：在超净工作台上吹干。 9.固定：于紫外交联仪内以1200 ENERGY照一次约半分钟，再重复一次，再在超净工作台上吹干