Selected article for: "PCR amplicon and Triton mm imidazole"
Author: Adedeji, Adeyemi O.; Marchand, Bruno; te Velthuis, Aartjan J. W.; Snijder, Eric J.; Weiss, Susan; Eoff, Robert L.; Singh, Kamalendra; Sarafianos, Stefan G.
Title: Mechanism of Nucleic Acid Unwinding by SARS-CoV Helicase
  • Document date: 2012_5_15
    • ID: 1ssh296a_32
    Snippet: Hexahistidine containing nsp13 was PCR amplified using the same primers mentioned above with BamHI and SalI restriction sites. The PCR amplicon was digested with BamHI and SacI, and ligated into pET-28a. A similar version of the plasmid was also obtained from Dr. John Ziebuhr (Justus Liebig University Giessen, Germany). Protein expression and purification was performed as described previously using Talon beads [35] . The protein was eluted with 2.....
    Document: Hexahistidine containing nsp13 was PCR amplified using the same primers mentioned above with BamHI and SalI restriction sites. The PCR amplicon was digested with BamHI and SacI, and ligated into pET-28a. A similar version of the plasmid was also obtained from Dr. John Ziebuhr (Justus Liebig University Giessen, Germany). Protein expression and purification was performed as described previously using Talon beads [35] . The protein was eluted with 25 mM Hepes pH 7.0, 0.5 M NaCl, 200 mM imidazole, 0.1% Triton X-100. It was further purified on a Superdex75 10/300GL column (20 mM Tris-HCl, pH 6.8, 200 mM NaCl, 1 mM DTT, and 5% glycerol. Fractions containing the desired protein were concentrated and stored at 280uC.

    Search related documents:
    Co phrase search for related documents
    • mm imidazole and protein expression: 1, 2, 3, 4, 5, 6, 7, 8, 9
    • mm imidazole and purification protein expression: 1, 2
    • mm imidazole and Triton mm imidazole: 1
    • PCR amplicon and protein expression: 1
    • PCR amplicon and purification protein expression: 1
    • PCR amplicon and Triton mm imidazole: 1
    • protein expression and purification protein expression: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25
    • protein expression and restriction site: 1, 2, 3
    • protein expression and Triton mm imidazole: 1
    • purification protein expression and Triton mm imidazole: 1