|Molecular Biology||Storage: -20°CShipping: Blue Ice|
5'-A T G C A^T-3'
3'-T^A C G T A-5'
Moraxella phenylpyruvica RFL1103
One unit is defined as the amount of Mph1103I required to digest 1ug of lambda DNA in 1 hour at 37°C in 50ul of assay buffer.
R1625: Restriction Enzyme Buffer A, 10X:
Dilute to 1X for use. 1X buffer composition is 33mM Tris-acetate pH 7.9, 10mM magnesium acetate, 66mM potassium acetate, 0.1mg/ml BSA.
R1625-04: Restriction Enzyme Buffer, 10X:
Dilute to 1X for use. 1X buffer composition is 10mM Tris-HCl, pH 8.5, 10mM MgCl2, 100mM potassium chloride,, 0.1mg/ml BSA
Supplied as a liquid in 10mM Tris-HCl (pH 7.4 at 25°C), 200mM KCl, 1mM DTT, 1mM EDTA, 0.2mg/ml BSA, 50% glycerol.
Stability during Prolonged Incubation: A minimum of 0.3 units of Mph1103I is required for complete digestion of 1ug of lambda DNA in 16 hours at 37°C.
Dam/Dcm/CpG/EcoKl: never overlaps- no effect
EcoBl: may overlap-effect not determined
Mph1103I is inactivated by incubation at 65°C for 20min.
Alw21I, PstI, SdaI, SduI
Number of Recognition Sites in DNA:
Storage and Stability:
May be stored at 4°C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20°C. Aliquots are stable for 6 months after receipt. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
Overdigestion Assay: No detectable change in the specific fragmentation pattern is observed after 160-fold overdigestion (10u/ug lambda DNA x 16 hours) with Mph1103I.
Ligation/Recutting Assay: The ligation and recleavage assay was replaced with LO test after validating experiments showed LO test ability to trace nuclease and phosphatase activities with sensitivity that is higher than L/R by a factor of 100.
Labeled Oligonucleotide (LO) Assay: No detectable degradation of a single-stranded and double-stranded labeled oligonucleotide was observed after incubation with 10 units of Mph1103I for 4 hours.
Blue/White Cloning Assay: The B/W assay was replaced with LO test after validating experiments showed LO test ability to detect nuclease and phosphatase activites with sensitivity that equals to that of B/W test.