RecAf |NEB酶试剂 New England Biolabs

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产品信息

产品类别:
Discontinued (<3 years)

  • 特性和用法

    反应条件

    1X Rec A Reaction Buffer
    Incubate at 37°C

    1X Rec A Reaction Buffer
    70 mM Tris-HCl
    10 mM MgCl2
    5 mM DTT
    (pH 7.6 @ 25°C)

    热失活

    65°C for 20 minutes

  • 优势和特性

    应用特性

    • Visualization of DNA structures with electron microscopy (3)
    • D-loop mutagenesis (4)
    • Screening libraries using RecA-coated probes (5,6)
    • Cleavage of DNA at any single predetermined site (7,8,9)
    • RecA mediated affinity capture for full length cDNA cloning (10,11)

  • 注意事项

    1. ATPγS which is required for triple helix formation is not supplied.
    2. Heat Inactivation: 65°C for 20 min

  • 参考文献

    1. West, S.C. (1992). Ann. Rev. Biochem. 61, 603-640.
    2. Zhumabayeva, B. et al. (1990). Biotechniques. 27, 834-845.
    3. Zhumabayeva, B. et al. (2001). Biotechniques. 30, 512-520.
    4. Pace, C.N. et al. (1995). Protein Sci. 4, 2411-2423.
    5. Radding, C.M. (1991). J. Biol. Chem. 266,
    6. Wasserman, S.A. and Cozzarelli, N.R. (1985). Proc. Natl. Acad. Sci. USA. 82, 1079-1083.
    7. Shortle, D. et al. (1980). Proc. Natl. Acad. Sci. USA. 77, 5375-5379.
    8. Honigberg, S.M. et al. (1986). Proc. Natl. Acad. Sci. USA. 83, 9586-9590.
    9. Rigas, B. et al. (1986). Proc. Natl. Acad. Sci. USA. 83, 9591-9595.
    10. Ferrin, L.J. and Camerini-Otero, R.D. (1991). Science. 254, 1494-1497.
    11. Koob, M. et al. (1992). Nucleic Acids Res. 20, 5831-5836.
    12. Koob, M. et al. (1992). In R. Wu(Ed.), Methods in Enzymology. 216, 321-329. San Diego: Academic Press.

操作说明、说明书 & 用法

  • 使用指南

    • DNA Damage and PreCR

FAQs & 问题解决指南

  • FAQs

    1. What buffer should be used with RecAf Protein?
    2. Is magnesium required for triple strand formation using RecAf Protein?
    3. Is ATP gamma-S required for triple strand formation using RecAf Protein?
    4. Can the RecAf Protein be heat inactivated?
    5. Is the structure of the target DNA important when using RecAf Protein?
    6. How much RecAf Protein should be added to the single stranded DNA to coat it?
    7. What length single stranded oligo should be used with RecAf Protein?

  • 实验技巧

    It’s N-term 6XHis tagged RecA with moderately stronger ssDNA binding ability compared to wt RecA. A possible reason might be the increased pI due to the attachment of the 6XHis-tag.