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M0250L |
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7,500 units
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10,000 units/ml |
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M0250S |
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1,500 units
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10,000 units/ml |
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- Removal of single-stranded extensions (3'and 5') to leave ligatable blunt ends
- Transcriptional mapping
- Cleavage of hairpin loops
- Supplied with 10X Reaction Buffer
Description:
A single-strand specific DNA and RNA endonuclease which will degrade single-stranded extensions from the ends of DNA and RNA molecules, leaving blunt, ligatable ends.
Source:
Mung bean sprouts
Applications:- Removal of 3“ and 5“ extensions from DNA or RNA termini
- Transcriptional mapping
- Cleavage of hairpin loops
- Excision of gene coding sequences from genomic DNA
- Generation of new restriction sites
Reagents Supplied:
Mung Bean Nuclease Reaction Buffer (10X)
Enzyme Properties
Molecular Weight:
Theoretical: 39 kDa
Reaction & Storage Conditions
Reaction Conditions:
1X Mung Bean Nuclease Reaction Buffer
Incubate at
30°C.
1X Mung Bean Nuclease Reaction Buffer:
50 mM sodium acetate
30 mM NaCl
1 mM ZnSO4
pH 5.0 @ 25°C
Unit Definition:
One unit is defined as the amount of enzyme required to produce 1 µg of acid-soluble total nucleotide in a total reaction volume of 50 μl in 1 minute at 37°C in 1X Mung Bean Nuclease Reaction Buffer with 0.5 mg/ml denatured Calf Thymus DNA.
Concentration:
10,000 units/ml
Storage Conditions:
10 mM sodium acetate
0.1 mM zinc acetate
1 mM Cysteine
50% Glycerol
0.001% Triton X-100
pH 5.0 @ 25°C
Storage Temperature:
-20°C
Notes
Usage notes:- It is no longer necessary to supplement Mung Bean Nuclease reactions with Zn2+. The zinc acetate in the storage buffer fulfills the Zn2+ requirement of the enzyme even after dilution in a reaction.
FAQs
- Should I use Mung Bean Nuclease or DNA Polymerase I, Large (Klenow) Fragment (NEB# M0210)?
- Is the Mung Bean Nuclease active in other NEBuffers?
- Can Mung Bean Nuclease be heat inactivated?
- Does Zn2+ need to be added to a Mung Bean Nuclease reaction?
- Will Mung Bean Nuclease degrade double stranded DNA?
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 double-strand exonuclease contamination.
Terminal Integrity:
After incubation of 1 μg of lambda DNA (Hae III digest) with 10 units of Mung Bean Nuclease
at 30ºC for 30 hour(s), > 95% of the DNA fragments can be ligated
with T4 DNA ligase. Of these ligated fragments, >95% can be recut with Hae III.
A) Molecular weight standard (λ HindIII digest)
B) M13mp8 (supercoiled)
C) As in B after incubation with NcoI
D) As in B after incubation with HindIII
E) As in D after incubation with Mung Bean Nuclease
F) As in E after incubation with T4 DNA Ligase
G) As in F after incubation with NcoI
H) As in F after incubation with HindIII
References
- Kowalski, D. et al. (1976) Biochemistry, 15, 4457-4463.
- McCutchan, T.F. et al. (1984) Science, 225, 626-628.
Reagents Sold Separately
Mung Bean Nuclease Reaction Buffer
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