标签归档：recaf

RecAf |NEB酶试剂 New England Biolabs

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上海金畔生物科技有限公司代理New England Biolabs（NEB）酶试剂全线产品，欢迎访问官网了解更多产品信息和订购。

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.

RecAf | NEB酶试剂 New England Biolabs

发表于由

上海金畔生物科技有限公司代理New England Biolabs（NEB）酶试剂全线产品，欢迎访问官网了解更多产品信息和订购。

RecAf

This product has been replaced with RecA (NEB #M0249).

RecAf is His-tagged. Native RecA is also available as a separate product.

  • Catalog # M0355 was discontinued on December 15, 2021
RecAf is a N-terminal 6X His tagged recombinant protein.  E. coli RecA is necessary for genetic recombination, reactions involving DNA repair and UV-induced mutagenesis. RecA promotes the autodigestion of the lexA repressor, umuD protein and lambda repressor. Cleavage of LexA derepresses more than 20 genes. In vitro studies indicate that in the presence of ATP, RecA promotes the strand exchange of single-strand DNA fragments with homologous duplex DNA. The reaction has three distinct steps: (i) RecA polymerizes on the single-strand DNA, (ii) the nucleo­protein filament binds the duplex DNA and searches for a homologous region, (iii) the strands are exchanged.