BLIRT TaqSSB – PCR Enhancer

PCR Enhancer

Name Part Number Pack Size
TaqSSB – PCR Enhancerl RP30 50 µg
  RP305 250 µg

 

Thermus aquaticus derived single-stranded DNA binding protein (SSB) is a thermostable protein which binds to single-stranded DNA with high specificity but does not bind well to double-stranded DNA (1). Thermus aquaticus SSB gene was expressed in E. coli in large quantities and the protein was highly purified.

Applications

- Protects against PCR inhibition (2)

- Enhances PCR yields (3,4,5,6,9)

- Enhances specificity and selectivity of multiplex PCR (7)

- Protects ssDNA from degradation by nucleases (8)

- Enables sequencing of complex-structure templates (i.e. hairpin)

- Enables amplification of GC-rich sequences

- Stimulates performance and fidelity of Taq polymerases (9)

- Stabilises ssDNA in site-specific mutagenesis

References

1. Greipel J., Urbanke C., and Maass G., Modulation of the affinity of the single-stranded DNA-binding protein of Escherichia coli (E. coli SSB) to poly(dT) by site-directed mutagenesis.  in: Saenger, W., Heinemann, U. (Eds.) pp. 61-86 (1989)

2. Kreader C., Relief of amplification inhibition in PCR with bovine serum albumin or T4 gene 32 protein. Applied Environ. Micro. 62, 1102-1106 (1996)

3. Dąbrowski S., Olszewski M., Piątek R. and Kur J., Novel thermostable ssDNA-binding proteins from Thermus thermophilus and T. aquaticus-expression and purification.  Protein Expr. Purif. 26, 131-138 (2002)

4. Dąbrowski S. and Kur J., Cloning, overexpression, and purification of the recombinant His-tagged SSB protein of Escherichia coli and use in polymerase chain reaction amplification.  Protein Expr. Purif. 16, 96-102 (1999)

5. Rapley R., Enhancing PCR amplification and sequencing using DNA-binding proteins. Mol. Biotech. 2, 295-298 (1994)

6. Schwarz K., Hansen-Hagge T. and Bartram C., Improved yields of long PCR products using gene 32 protein. Nucleic Acids Res. 18, 1079 (1990)

7. Barski P., Piechowicz L., Galinski J. and Kur J., Rapid assay for detection of methicillin-resistant Staphylococcus aureus using multiplex PCR. Mol. Cell Probes 10, 471-475 (1996)

8. Fradkin G, Aizenberg O, Torosian M, Shishkova O, Enzymatic mechanisms of degradation of DNA replication forks in vitro. Mol Biol. 18, 1518-23 (1984)

9. Perales C., Cava F., Meijer W., Berenguer J., Enhancement of DNA, cDNA synthesis and fidelity at high temperatures by a dimeric single-stranded DNA-binding protein.  Nucleic Acids Res. 31, 6473-6480 (2003)

Source

Thermus aquaticus

Storage temperature

-20°C

Storage Conditions

10 mM Tris-HCl (pH 7.5 at 22°C), 50 mM NaCl, 0.1 mM DTT, 0.05% Triton X-100, 0.1 mM EDTA and 50% (v/v) glycerol.

Quality Control

Greater than 95% of protein determined by SDS-PAGE (CBB staining). The absence of endonucleases and exonucleases was confirmed.

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