标签: Phenol-based
顶[0] 发表评论(5) 编辑词条
Phenol-based Method for the Isolation of DNA Fragments from Low-Melting Temperature Agarose Reference: Favre, D. 1992. Biotechniques vol. 13 1.Cut out slice containing DNA, smallest size possible. 2.Estimate volume and double with TE (10 mM Tris-HCl, pH 8.0/1 mM EDTA). Melt at 65 ℃ 5-10 min. 3.Add 1 volume Tris-buffered phenol at room temperature, mix by inversion. 4.Spin 3 min at 10-12k rpm and transfer aqueous phase to new tube. Phenol extract again. 5.Spin as in (4) and transfer aqueous phase to new tube containing 0.1 volume 4 M LiCl. Mix by inversion; a white precipitate forms immediately. Place tube on ice 2 min. Spin as above, 3 min. 6.Transfer to new tube, leaving transparent pellet behind. Add 1 μl carrier (glycogen) and precipitate with 2.5 volumes cold ethanol. Mix, leave at -70 ℃ 5-10 min, and spin as above, 10 min. Wash pellet with 1 ml 70% ethanol, dry under vacuum, and resuspend in 10-20 μl water or TE. |
→如果您认为本词条还有待完善,请 编辑词条
上一篇操作DNA(第三部分) 下一篇什么是克隆技术
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