| 查看: 275 | 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
售一区SCI文章T0P，我:8O.551.O54,科目全,可十急
已经有4人回复
-
师弟论文见刊大半年才想起来申请专利，还能抢救一下吗？
已经有5人回复
-
某top大学教授说“能够在市场中兑现的能力才是真能力”无比同意！
已经有3人回复
-
售SCI文章，我:8O.5.5.1O.54,科目全,可十急
已经有3人回复
-
售SCI一区T0P文章，我:8O.55.1.O.54,科目全,可伽急
已经有3人回复
-
售SCI一区T0P文章，我:8O.55.1.O.5.4,科目齐全,可+急
已经有3人回复
-
售SCI一区T0P文章，我:8.O.55.1.O54,科目全,可伽急
已经有3人回复
-
售SCI文章，我:8O5.5.1.O.54,科目齐全,可+急
已经有3人回复
-
售SCI一区T0P文章，我:8.O.55.1.O.54,科目齐全,可+急
已经有3人回复
-
售SCI文章，我:8O5.5.1.O.54,科目齐全,可+急
已经有4人回复
2楼2009-04-28 20:48:12
3楼2009-05-05 09:03:33
