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DNA Modifying Enzymes and Cloning >
Single-stranded DNA Binding Proteins >
T4 Gene 32 Protein |
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Prices are in US dollars and valid only for US orders.
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- Stabilize and mark ssDNA for elctron microscopy
- Improve yield and efficiency of RT during RT-PCR
- Isolated from a recombinant source
Description:
T4 Gene 32 Protein is a single-stranded DNA (ssDNA) binding protein required for bacteriophage T4 replication and repair (1). It cooperatively binds to and stablizes transiently formed regions of ssDNA and plays an important structural role during T4 phage replication (2). It also has been used extensively to stabilize and mark regions of ssDNA for electron microscopic examination of intracellular DNA structures (3). Recently, it has been shown to improve the yield and efficiency of reverse transcription (RT) reactions during RT-PCR as well as increase the yield of PCR products amplified from humic acid containing soil samples (4-7).
Source:
A E. coli strain carrying a plasmid that overexpresses the gene 32 protein of T4 phage
Applications:- Stabilization and marking of ssDNA structures (3)
- Increase yield and specificity of PCR products from soil samples (4)
- Increase yield and processivity of RT during RT-PCR (5-7)
Reagents Supplied:
NEBuffer 4
Properties & Usage
Heat Inactivation:
65°C for 20 minutes
Molecular Weight:
Theoretical: 33,485 daltons
Reaction & Storage Conditions
Reaction Conditions:
1X NEBuffer 4
Incubate at
37°C.
1X NEBuffer 4:
20 mM Tris-acetate
50 mM potassium acetate
10 mM Magnesium Acetate
1 mM Dithiothreitol
pH 7.9 @ 25°C
Unit Definition:
Sold by mass of pure protein as determined by OD280 (A280 = 0.516 at 1 mg/ml, 1 cm)
Concentration:
10 mg/ml
Storage Conditions:
20 mM Tris-HCl
100 mM NaCl
0.5 mM Dithiothreitol
1 mM EDTA
50% Glycerol
pH 8.0 @ 25°C
Storage Temperature:
-20°C
Quality Control for Current Lot
Quality control values for a specific lot can be found on the datacard which accompanies each product.
Quality Assurance Statement:
Purified free of contaminating endonucleases and exonucleases. Each lot is tested for ssDNA binding by its ability to produce a mobility-shift of 2.5 μg ΦX174 virion DNA (single-stranded) using 10 μg T4 Gene 32 Protein as the standard.
References
- Alberts, B. and Frey, L. (1970) Nature, 227, 1313-1318.
- Bittner, M. et al. (1979) J. Biol. Chem., 254, 9565-9572.
- Delius, H. et al. (1972) J. Mol. Biol., 67, 341-350.
- Tebbe, C.C. and Vehjen, W. (1993) Appl. Environ. Microbiol.
, 59, 2657-2665.
- Villalva, C. et al. (2001) Biotechniques, 31, 81-86.
- Baugh, L. R. et al. (2001) Nucl. Acids Res., 29, e29.
- Jeffries, D. and Farquharson, C. (2002) Anal. Biochem., 308, 192-194.
Reagents Sold Separately
NEBuffer 4
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