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. 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. 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

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.

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.

Interactive Tools

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.

Safety Data Sheet

The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.

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.