Procedure Add the following to a sterile microcentrifuge tube: Labeled target DNA (3-6ng) 1-8ul Calf Thymus DNA (0.5ug/ul) 2ul TE buffer 0-7ul Total Volume 10ul Add 1ul of 4% Formic Acid and incubate for 25 min at 37℃ During this incubation, add 15 ul of stock piperidine to 135 ul of water to prepare a 1M piperidine solution. Place the tube containing the formic acid reaction on ice, add all 150 ul of the diluted (1M) piperidine solution and incubate for 30 min at 90℃. Place this reaction on ice for 5min, add 1ml of n-butanol and vortex. Centrifuge for 2min at high speed to pellet the DNA. Remove the supernatant and add 150ul of 1% SDS to the pellet. Add 1ml of n-butanol, vortex vigorously and centrifuge at high speed for 2 min. Carefully remove the supernatant. Add 0.5 ml of n-butanol to rinse the pellet, and carefully remove the supernatant. Repeat this rinse step once.
Note This protocol was adopted from Amersham footprinting kit instruction. |
→如果您认为本词条还有待完善,请 编辑词条
上一篇粒度仪器选购指南 下一篇蛋白质组学研究中的核心技术-双向凝胶电泳
词条内容仅供参考,如果您需要解决具体问题
(尤其在法律、医学等领域),建议您咨询相关领域专业人士。
0