|Molecular Biology||Storage: -20°CShipping: Blue Ice|
5'-C^A A T T G-3'
3'-G T T A A^C-5'
Source: E. coli that carries the cloned munIR gene from Mycoplasma unidentified.
Storage Buffer: 10mM Tris-HCl (pH 7.4 at 25°C), 100mM KCl, 1mM DTT, 1mM EDTA, 0.2mg/ml BSA and 50% glycerol.
Diluent Buffer: 10mM Tris-HCl (pH 7.4 at 25°C), 100mM KCl, 1mM EDTA, 1mM DTT, 0.2mg/ml BSA and 50% glycerol.
R1625-Restriction Enzyme Buffer A, 10X:
Dilute to 1X for use. 1X buffer composition is 33mM Tris-acetate (pH 7.9 at 37°C), 10mM magnesium acetate, 66mM potassium acetate, 0.1mg/ml BSA.
R1625-02-Restriction Enzyme Buffer C, 10X:
Dilute to 1X for 100% MunI digestion. 1X buffer compostion is 10mM Tris-HCl (pH 7.5), 10mM MgCl2, 50mM NaCl and 0.1mg/ml BSA. Incubate at 37°C
Storage and Stability:
May 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.
Overdigestion Assay: No detectable change in the specific fragmentation pattern is observed after 160-fold overdigestion (10u/ug lambda DNA x 16 hours) with MunI.
Ligation/Recutting Assay: After 50-fold overdigestion (3u/ug DNA x 17 hours) with MunI, more than 95% of the DNA fragments can be ligated at a 5'-termini concentration of 0.07uM. 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 10 units of enzyme for 4 hours.
Blue/White Cloning Assay: The mix of pUC57/HindIII, pUC57/PstI and pUC57/Eco32I digests was incubated with 10 units of enzyme for 16 hours. After religation and transformation < 1% of white colonies were detected.
Stability during Prolonged Incubation: A minimum of 0.1 units of enzyme is 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.
Digestion of Agarose-embedded DNA: A minimum of 5 units of enzyme is required for digestion of 1ug of agarose-embedded lambda DNA in 16 hours.
XapI, EcoRI, TasI
Number of Recognition Sites in DNA:
Methylation Effect on Digestion:
Dam, Dcm, CpG, EcoKI: Never overlaps - no effect.
EcoBI: May overlap - effect undetermined.
Unit Definition: 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.
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.