Selected article for: "lysis buffer and Tris buffer"
Author: Shu, Ting; Gan, Tianyu; Bai, Peng; Wang, Xiaotong; Qian, Qi; Zhou, Hui; Cheng, Qi; Qiu, Yang; Yin, Lei; Zhong, Jin; Zhou, Xi
Title: Ebola virus VP35 has novel NTPase and helicase-like activities
  • Document date: 2019_6_20
    • ID: u3pxycqh_30
    Snippet: RNA-IP was performed as previously described (42) with minor modification. Briefly, cells were lysed in a lysis buffer [20 mM Tris-HCl (pH 7.4), 200 mM NaCl, 2.5 mM MgCl 2 , 0.5% Triton X-100, 0.5 U/ml RNase inhibitor (Promega) and a protease inhibitor cocktail (Roche)] at 4 • C for 30 min. Lysates were clarified at 12 000 g for 10 min at 4 • C and the post-nuclear lysates were pre-cleared by incubation with protein-A/G agarose beads (Roche) .....
    Document: RNA-IP was performed as previously described (42) with minor modification. Briefly, cells were lysed in a lysis buffer [20 mM Tris-HCl (pH 7.4), 200 mM NaCl, 2.5 mM MgCl 2 , 0.5% Triton X-100, 0.5 U/ml RNase inhibitor (Promega) and a protease inhibitor cocktail (Roche)] at 4 • C for 30 min. Lysates were clarified at 12 000 g for 10 min at 4 • C and the post-nuclear lysates were pre-cleared by incubation with protein-A/G agarose beads (Roche) at 4 • C for 2 h. Then the pre-cleared lysates were incubated with antibodies (anti-Flag or anti-IgG) together with protein-A/G agarose beads (Roche) at 4 • C for 12 h. The antibody-bound complexes were washed for five times with the same lysis buffer. Finally, proteins or RNAs were extracted from the complexes and analyzed by northern or western blotting as describe above.

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