| 查看: 241 | 回复: 2 | |||
| 当前主题已经存档。 | |||
[交流]
请问northern blot杂交液只用7% SDS in 0.25 M sodium phosphate buffer pH 7.2行吗
|
|||
|
大家好 我看有些文献上做siRNA BLOT 只用7% SDS in 0.25 M sodium phosphate buffer pH 7.2 请问有人做过吗,谢谢 如: Northern blot analyses Total RNA (30–50 g) was separated on a 15% denaturing polyacrylamide Tris/borate/EDTA gel; the gel was then soaked in 10 mM sodium phosphate buffer pH 7.0 and subsequently in 20 SSC for 10 min. The RNAs were transferred by overnight capillary blotting in 20 SSC to Zeta-probe (Bio-Rad) or Hybond N+ (Amersham Biosciences) membranes. Membranes were crosslinked and/or baked for 2 h at 80 °C. Probes complementary to sRNAs were end-labelled with [ -32P]ATP with the use of T4 polynucleotide kinase (New England BioLabs). Hybridization was performed in 5 ml of 7% SDS in 0.25 M sodium phosphate buffer pH 7.2, and the hybridization temperature was set 15 °C below the Tm of oligonucleotides. Membranes were washed twice at hybridization temperature with 2 SSC, 0.1% SDS for 10 min, and exposed to PhosphorImage plates. Decade RNA marker was labelled in accordance with the manufacturer's instruction (Ambion). |
回复此楼» 猜你喜欢
售SCI一区T0P文章，我:8O.55.1.O.5.4,科目齐全,可+急
已经有4人回复
-
今天系统多次维护，明天很可能放榜！
已经有7人回复
-
哪位老哥知道今年的国自然具体哪一天放榜？
已经有10人回复
-
售SCI一区T0P文章，我:8.O.55.1.O.5.4,科目全,可+急
已经有3人回复
-
欢迎发来filecode的Mz6后的代码验证其规律
已经有74人回复
-
岩土工程学报什么时候才能终审完呐
已经有4人回复
-
时间戳又变了8-15
已经有25人回复
-
2027广东省杰青
已经有6人回复
-
快农历七夕节了，轻松一下，男人悄悄话，女施主请不要进来。
已经有3人回复
-
咱们一起用铁证分析2026国家社科基金中标与否
已经有26人回复
2楼2009-04-28 20:48:12
3楼2009-05-05 09:03:33
