|Molecular Biology||Storage: -20°CShipping: Blue Ice|
5'...C C T C (N)7 ...3'
3'...G G A G (N)6 ...5'
E. coli with cloned mnlIR gene from Moraxells nonliquefaciens
One unit is defined as the amount of enzyme required to digest 1ug of lambda DNA in 1 hour at 37°C in 50ul of assay buffer.
10mM Tris-HCl, pH 7.4 at 25°C, 100mM KCl, 1mM DTT, 1mM EDTA, 0.2mg/ml BSA, 50% glycerol.
Dilute in the same buffer as Form above.
R1625: Restriction Enzyme Buffer A, 10X
R1625-02: Restriction Enzyme Buffer C, 10X
Note: R1625-02 gives 100% digestion where R1625 will only give ~20-50% digestion at the same concentrati
Storage and Stability:
Aliquot to avoid repeated freezing and thawing. 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.
Dam: Never overlaps - no effect
Dcm: Never overlaps - no effect
CpG: May overlap - no effect
EcoKI: Never overlaps - no effect
EcoBI: May overlap - Blocked
Stability during Prolonged Incubation: A minimum of 0.5units of enzyme are required for complete digestion of 1ug of lambda DNA in 16 hours at 37°C.
Thermal Inactivation: Enzyme is inactivated by incubation at 65°C for 20min.
Number of Recognition Sites in DNA:
• Mnl I produces DNA fragments that have a single base 3’-extension which are more difficult to ligate than blunt-ended fragments.
• Mnl I may remain associated with the cleaved DNA. This may cause DNA band shifting during electrophoresis. To avoid atypical DNA band patterns, heat the digested DNA in the presence of SDS prior to electrophoresis.
Overdigestion Assay: No detectable change in the specific fragmentation pattern is observed after 160-fold overdigestion (10u/ug lambda DNA x 16 hours) with Mnl I.
Ligation/Recutting Assay: After 50-fold overdigestion (3u/ug DNA x 17 hours) with Mnl I, more than 90% of the DNA fragments can be ligated at a 5'-termini concentration of 2uM. More than 90% 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 restriction endonuclease for 4 hours.
Important Note: This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications without the expressed written authorization of United States Biological.