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  • ET SSB

    Description

    ET SSB (Extreme Thermostable Single-Stranded DNA Binding Protein) is a single-stranded DNA binding protein isolated from a hyperthermophilic microorganism, which remains fully active after incubation at 95°C for 60 minutes. Due to the extreme thermostability, ET SSB can be used in applications that require extremely high temperature conditions, such as nucleic acid amplification and sequencing.

    Figure 1: Improved multiplex PCR by ET SSB.
    By adding ET SSB, specific amplification was accomplished in PCR using one to five primer pairs in incurring order (lanes 1-5). Lane 5C shows a PCR reaction using 5 primer pairs, with no ET SSB included (control reaction). Lane M: 100 bp DNA Ladder.
    Figure 2: Multiplex HDA was improved by ET SSB.
    The HDA reactions were prepared with either single or two sets of primers to amplify the invE (A) and/or invA (B) from Salmonella typhimurium genomic DNA. Lane M: Low Molecular DNA Ladder; Lane A: invE gene; Lane B: invA gene; Lane AB: invE and invA genes with no ET SSB present; Lane AB**: invE and invA genes with ET SSB in the HDA reaction. ** the reaction contained ET SSB.

    Product Source

    An E. coli strain that carries the cloned ssb gene from a hyperthermophilic organism

    Properties and Usage

    Unit Definition

    Sold by mass of pure protein as determined by OD280.

    Storage Temperature

    -20°C

    Storage Conditions

    20 mM Tris-HCl
    200 mM NaCl
    0.5 mM DTT
    1 mM EDTA
    50% Glycerol
    pH 7.5 @ 25°C

    Heat Inactivation

    No

    Quality Control

    Quality Assurance Statement

    • ET SSB is purified free of contaminating endonucleases and exonucleases. Each lot is tested for ssDNA binding activity and is visually determined to be > 95% pure on an SDS-polyacrylamide gel.

    Notes

    1. ET SSB is active in any polymerase buffer. Add 200 ng of ET SSB per 50 µl reaction.

    References

    1. Myers, T.W. and Romano, L.J. (1988). J. Biol. Chem. 263, 17006-17015.
    2. Delius, H., Mantell, N.J. and Alberts, B. (1972). J. Mol. Biol.. 67, 341-350.
    3. Schwarz, K., Hansen-Hagge, T. and Bartram, C. (1990). Nucl. Acids Res. 18, 1079.
    4. Chou, Q. (1992). Nucl. Acids Res. . 20, 4371.
    5. Oshima, R.G. (1992). Biotechniques. 13, 188.
    6. Rapley, R. (1994). Mol. Biotechnol.. 2, 295-298.
    7. Olszewski, M. et al. (2005). Mol. Cell Probes. 19, 203-205.
    8. Baugh, L.R. et al. (2001). Nucl. Acids Res.. 29, e29.
    9. Villalva, C. et al. (2001). Biotechniques. 31, 81-83, 86.
    10. Reddy, M.S. (2000). Biochemistry. 39, 14250-14262.
    11. West, S.C., Cassuto, E. and Howard-Flanders, P. (1982). Mol. Gen. Genet.. 186, 333-338.
    12. Goldmeyer, J., Kong, H., and Tang, W. (2007). J. Mol. Diagnostics. 9, 639-644.

    Supporting Documents

    Material Safety Datasheets

    The following is a list of Material Safety Data Sheets (MSDS) that apply to this product to help you use it safely. The following file naming structure is used to name these document files: [Product Name] MSDS. For international versions please contact us at info@neb.com.

    Datacards

    The Product Summary Sheet, or Data Card, includes details for how to use the product, as well as details of its formulation and quality controls. The following file naming structure is used to name the majority of these document files: [Catalog Number]Datasheet-Lot[Lot Number]. For those product lots not listed below, please contact NEB at info@neb.com or fill out the Technical Support Form for appropriate document.