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DNA Modifying Enzymes and Cloning >
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T7 Exonuclease |
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Prices are in US dollars and valid only for US orders.
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- Isolated from a recombinant source
- 5' → 3' exonuclease
- Supplied with 10X Reaction Buffer
Description:
T7 Exonuclease acts in the 5' to 3' direction, catalyzing the removal of 5' mononucleotides from duplex DNA. T7 Exonuclease is able to initiate nucleotide removal from the 5' termini or at gaps and nicks of double-stranded DNA (1). It will degrade both 5' phosphorylated or 5' dephosphorylated DNA. It has also been reported to degrade RNA and DNA from RNA/DNA hybrids in the 5' to 3' direction but is unable to degrade either double-stranded or single-stranded RNA (2). The protein is the product of T7 gene 6.
Source:
Purified from an E. coli strain containing a TYB12 intein fusion.
Reagents Supplied:
NEBuffer 4 (10X)
Enzyme Properties
Heat Inactivation:
No
Reaction & Storage Conditions
Reaction Conditions:
1X NEBuffer 4
Incubate at
25°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:
One unit is defined as the amount of enzyme required to produce 1 nmol of acid-soluble deoxyribonucleotide from double-stranded DNA in a total reaction volume of 50 μl in 30 minutes at 25°C in 1X NEBuffer 4 with 0.15 mM sonicated duplex [3H]-DNA.
Concentration:
10,000 units/ml
Storage Conditions:
10 mM Tris-HCl
5 mM Dithiothreitol
0.1 mM EDTA
50% Glycerol
pH 8.0 @ 25°C
Storage Temperature:
-20°C
FAQs
- How Does T7 Exonuclease differ from Lambda Exonuclease (NEB# M0262)?
- How Does T7 Exonuclease differ from Exonuclease III (NEB# M0206)?
- What is the activity of T7 Exonuclease in the NEBuffers?
- Can T7 Exonuclease be heat inactivated?
- What is the best way to generate partials using T7 Exonuclease?
- Can DNA be blunted using T7 Exonuclease?
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.
Endonuclease Activity:
Incubation of a 50 μl reaction containing 400 units of T7 Exonuclease with 1 μg of
ΦX174 RF I DNA for 4 hours at 25ºC resulted
in < 10% conversion to RFII as determined by agarose gel electrophoresis.
RNase Assay Radioactive:
Incubation of 10 units of enzyme with 20 nmol [3H]poly(A).poly(dT) hybrid polymer for
1 hour at 37ºC in a 50 μl reaction released < 15.2
nmol adenosine-5'-monophosphate.
SS DNA Exonuclease Activity:
Incubation of 10 units of enzyme with 1 μg sonicated and denatured [3H]-DNA (105 cpm/μg) for 30 minutes at 25ºC in 50 μl reaction buffer released < 1.5% radioactivity.
References
- Kerr, C. and Sadowski, P.D. (1972) J. Biol. Chem., 247, 305-318.
- Shinozaki, K. and Okazaki, T. (1978) Nucl. Acids Res., 5, 4245-4261.
Reagents Sold Separately
NEBuffer 4
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