ET SSB can be used in applications that require extremely high temperature conditions, such as nucleic acid amplification and sequencing.
Improve the processivity of DNA polymerase
Stabilization and marking of ssDNA structure
Increase the yield and specificitiy of PCR
Increase the yield and processivity of RT during RT-PCR
Improve DNA sequencing through regions with strong secondary structure
Product Information
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
This product is related to the following categories:
The following reagents are supplied with this product:
NEB #
Component Name
Component #
Stored at (°C)
Amount
Concentration
M2401S
-20
ET SSB
M2401SVIAL
-20
1 x 0.1 ml
500 µg/ml
Properties & Usage
Unit Definition
Sold by mass of pure protein as determined by OD280.
Storage Buffer
10 mM Tris-HCl
100 mM KCl
0.1 mM EDTA
50% Glycerol
pH 7.4 @ 25°C
Heat Inactivation
No
Molecular Weight
Apparent: 16 kDa
Product Notes
ET SSB is active in any polymerase buffer. Add 200 ng of ET SSB per 50 µl reaction.
References
Myers, T.W. and Romano, L.J. (1988). J. Biol. Chem. 263, 17006-17015.
Reddy, M.S. (2000). Biochemistry. 39, 14250-14262.
West, S.C., Cassuto, E. and Howard-Flanders, P. (1982). Mol. Gen. Genet. 186, 333-338.
Goldmeyer, J., Kong, H., and Tang, W. (2007). J. Mol. Diagnostics. 9, 639-644.
Delius, H., Mantell, N.J. and Alberts, B. (1972). J. Mol. Biol. 67, 341-350.
Schwarz, K., Hansen-Hagge, T. and Bartram, C. (1990). Nucl. Acids Res. 18, 1079.
Chou, Q. (1992). Nucl. Acids Res. 20, 4371.
Oshima, R.G. (1992). Biotechniques. 13, 188.
Rapley, R. (1994). Mol. Biotechnol. 2, 295-298.
Olszewski, M. et al. (2005). Mol. Cell Probes. 19, 203-205.
Baugh, L.R. et al. (2001). Nucl. Acids Res. 29, e29.
Villalva, C. et al. (2001). Biotechniques. 31, 81-83, 86.
Citations & Technical Literature
Citations
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