Single-strand binding protein
- Enhances DNA polymerase proofreading activity and PCR fidelity
- Increases yield of large PCR DNA fragments
T4 bacteriophage gene 32 product, or T4 gp32, is a non-sequence specific, single-strand binding protein isolated from the bacteriophage T4. It is reported to act by binding to single-stranded DNA in a co-operative manner thereby destabilizing double-stranded DNA (1).
Used with either Taq or Arrow Taq DNA polymerase, T4 gp32 protein strongly increases PCR fidelity. The T4 gp32 protein is used to improve yields of long PCR products catalyzed by Taq DNA Polymerase (2), DNA polymerase proofreading (3) and amplification efficiency in low purity DNA. It has also been used to stabilize single-stranded regions of DNA for site-directed mutagenesis.
|Storage buffer:||Tris-HCl 20 mM, pH 8.0, NaCl 100 mM, EDTA 1 mM, DTT 0.5 mM, Glycerol 50%.|
The supplied protein is a purified recombinant form free of endonucleases and nickases.
Improved yield of long PCR product (25.7 kb) in the presence of T4 gp32 DNA template was amplified with Arrow Taq.
Reaction conditions: 50µL reaction; 10 ng λ DNA, 50 pmol each primer, 300 µM each dNTP, 5 U Arrow Taq with or without T4gp32 in 1 x Tpol buffer (1.5 mM MgCl2), 5 sec 94°C, 2 min at 65°C, 30 min at 72°C, 20 cycles.
Lane 1: 5U Arrow Taq without T4 gp32; Lane
2: 5U Arrow Taq with 10 µg/ml T4gp32; Lane
3: λ/EcoO 109 I
1. Topal, M.D. and Sinha, N.K. (1983). Products of bacteriophage T4 genes 32 and 45 improve the accuracy of DNA replication in vitro. J. Biol. Chem. 258: 12274-12279.
2. Schwarz, K., Hansen-Hagge, T., Bartram, C. (1990). Improved yields of long PCR products using gene 32 protein. Nucleic Acids Research 18 :1079.
3. Sandhu, D.K., Keohavong, P. (1994). Effects of the T4 bacteriophage gene 32 product on the efficiency and fidelity of DNA amplification using T4 DNA polymerase. Gene 144: 53-58.