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DNA Modifying Enzymes and Cloning >
Nucleases >
Lambda 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
- Highly processive 5'→3' exonuclease
- Supplied in 10X Reaction Buffer
Description:
A highly processive enzyme that acts in the 5´ to 3´ direction, catalyzing the removal of 5´ mononucleotides from duplex DNA. The preferred substrate is 5´-phosphorylated double stranded DNA, although it will also degrade single-stranded and non-phosphorylated substrates at a greatly reduced rate. Lambda Exonuclease is unable to initiate DNA digestion at nicks or gaps (1).
Source:
A genetic fusion of the E. coli Lambda Exonuclease gene with the gene encoding maltose binding protein (MBP). Following affinity chromatography, Lambda Exonuclease is cleaved from the fusion construct and purified away from MBP.
Reagents Supplied:
Lambda Exonuclease Reaction Buffer (10X)
Enzyme Properties
Heat Inactivation:
75°C for 10 minutes
Molecular Weight:
Theoretical: 28,000 daltons
Specific Activity:
100,000 units/mg
Reaction & Storage Conditions
Reaction Conditions:
1X Lambda Exonuclease Reaction Buffer
Incubate at
37°C.
1X Lambda Exonuclease Reaction Buffer:
67 mM Glycine-KOH
2.5 mM MgCl2
50 µg/ml BSA
pH 9.4 @ 25°C
Unit Definition:
One unit is defined as the amount of enzyme required to produce 10 nmol of acid-soluble deoxyribonucleotide from double-stranded substrate in a total reaction volume of 50 µl in 30 minutes at 37°C in 1X Lambda Exonuclease Reaction Buffer with 1 µg sonicated duplex [3H]-DNA.
Concentration:
5,000 units/ml
Storage Conditions:
25 mM Tris-HCl
50 mM NaCl
1 mM Dithiothreitol
0.1 mM EDTA
50 µg/ml BSA
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.
Endonuclease Activity:
Incubation of a 50 μl reaction containing 200 units of Lambda Exonuclease with 1 μg of
ΦX174 RF I DNA for 4 hours at 37ºC resulted
in < 10% conversion to RFII as determined by agarose gel electrophoresis.
References
- Little, J.W. (1981) Gene Amplif. Anal., 2, 135-145.
Reagents Sold Separately
Lambda Exonuclease Reaction Buffer
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