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You are here:Home » Molecular Biology » MB-Enzymes, Restriction » Alw26I (BsmAI)

Alw26I (BsmAI)

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Specifications

5'-G T C T C (N)1^-3'
3'-C A G A G (N)5^-5'
Catalog #A1372-10
SourceAcinetobacter lwoffi RFL26
Concentration10u/ul
Unit DefinitionOne unit is defined as the amount of Alw26I required to digest 1ug of lambda DNA in 1 hour at 37°C in 50ul of assay buffer.
Incubation Temperature37°C
Diluent Buffer10mM Tris-HCl, pH 7.4, 100mM KCl, 1mM EDTA, 1mM DTT, 0.2mg/ml BSA and 50% glycerol.
Storage BufferSupplied as a liquid in 10mM Tris-HCl, pH 7.4, 100mM KCl, 1mM DTT, 1mM EDTA, 0.2mg/ml BSA and 50% glycerol.
Restriction Enzyme Buffer A, 10XR1625: Supplied as a liquid in 33mM Tris-acetate, pH 7.9, 10mM magnesium acetate, 66mM potassium acetate and 0.1mg/ml BSA. Incubate at 37°C. Supplied to simplify buffer selection for double digests.
Storage and StabilityMay be stored at 4°C for short-term only. For long-term storage, aliquot and store at -20°C. Aliquots are stable for at least 6 months at -20°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.
Quality Control
Overdigestion Assay
No detectable change in the specific fragmentation pattern is observed after 160-fold overdigestion (10u/ug lambda DNA x 16 hours) with Alw26I.
Ligation/Recutting Assay
After 50-fold overdigestion (3u/ug DNA x 17 hours) with Alw26I more than 95% of the DNA fragments can be ligated at a 5'-termini concentration of 0.1uM. More than 95% of these can be recut.
Labeled Oligonucleotide (LO) Assay
No detectable degradation of a single-stranded and double-stranded labeled oligonucleotide was observed after incubation with 10units of Alw26I for 4 hours.
Methylation Effects
Alw26I does not cut GAGAm5C. Overlapping CpG methylation may influence DNA cleavage.
Stability during Prolonged Incubation
A minimum of 0.2units of Alw26I is required for complete digestion of 1ug of lambda DNA in 16 hours at 37°C.
Thermal Inactivation
Alw26I is inactivated by incubation at 65°C for 20min.
Number of Recognition Sites in DNA
Lambda37
PhiX1744
M13mp18/195
pBR3223
pUC18/194
pUC574
pTZ19R/U2
Protocol for Digestion
Add
Nuclease free water16ul
R16252ul
DNA (0.5-1ug/ml)1ul
A1372-100.5-2ul
Mix gently and spin down for a few seconds. Incubate at 37ºC for 1-16 hours.
Protocol for Digestion Directly after Amplification
Add
PCR Reaction Mixture10ul (~0.1-0.5ug of DNA)
Nuclease free water18ul
R16252ul
A1372-101-2ul
Mix gently and spin down for a few seconds.
Incubate at 37ºC for 1-16 hours.


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