1。杂交过程一个杂交管会采取1或2的膜(22厘米大小22)。使用杂交液50毫升和0.5毫克每杂交管的DNA探针(10 ng / ml的终浓度)。为了达到最佳效果,请使用凝胶纯化的DNA为探针。如果缩放了探针的制备,管数量的增加,而不是增加每管中的试剂量。 探针制备(0.5毫克DNA): 60μL水稀释15μL交联剂的解决方案。 稀的DNA为10 ng /μL水,一个螺丝帽?…
Contains labeling reagent, cross-linker solution, reaction buffer, water, control DNA, hybridization buffer, blocking reagent. Store hybridization buffer and blocking reagent at room temperature, other reagents at 2-8°C.
ECF Substrate
kit
Amersham
RPN3685
Contains ECF substrate, ECF substrate dilution buffer.
Note: Pack contains total 60 ml reagent; aliquot into 10 ml samples in Falcon tube and store at 20°C. Use about 10 ml per membrane (size of 22 by 22 cm).
20x Secondary Wash Buffer
buffer
121 g Tris (final 1 M),
117 g NaCl (final 2 M).
Adjust pH to 10. Adjust to 1 L with water. Store at 2-8°C for up to 4 months.
0.5 M NaH2PO4 buffer pH 7
buffer
69 g NaH2PO4.H2O.
Adjust pH to 7 with NaOH. Adjust to 1 L with water. Store at room temperature.
1 M MgCl2
50.8 g MgCl2.6H2O
Adjust to 250 ml with water. Store at room temperature.
10% (w/v) SDS
20 g SDS
Adjust to 200 ml with water. Store at room temperatur