5'-C ↓ Y C G R G 3'
3'-G R G C Y ↑C 5'

Source:
E. coli that carries the cloned Eco88IR gene from E. coli RFL88

Unit Definition:
One unit is defined as the amount of Eco88I required to digest 1ug of lambda DNA in 1 hour at 37°C in 50ul of assay buffer.

Storage Buffer:
10mM Tris-HCl, pH 7.4, 100mM KCl, 1mM EDTA, 1mM DTT, 0.2mg/ml BSA and 50% glycerol

Supplied with:
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

Incubation Temperature:
37ºC

Thermal Inactivation:
Eco88I is inactivated by incubation at 65°C for 20 minutes

Diluent Buffer:
10mM Tris-HCl, pH 7.4, 100mM KCl, 1mM EDTA, 1mM DTT, 0.2mg/ml BSA and 50% glycerol

Enzyme Properties:
Methylation Effects:
Dam, Never overlaps - no effect
Dcm: Never overlaps - no effect
CpG: Completely overlaps - cleavage impaired
EcoKI: Never overlaps - no effect
EcoBI: Never overlaps - no effect

Stability during Prolonged Incubation:
A minimum of 0.2units of Eco88I is required for complete digestion of 1ug of lambda DNA in 16 hours at 37°C.

Digestion of Agarose-embedded DNA:
A minimum of 5 units of Eco88I is required for complete digestion of 1ug of agarose-embedded lambda DNA in 16 hours.

Compatible Ends:
C↓CCGGG - BshTI, BsaWI, Cfr9I, Cfr10l, Kpn2l, Mrel, NgoMIV, SgAI

C↓TCGAG - Sall, SgrDI, Smol, Xhol

Number of Recognition Sites in DNA:
Lambda: 8
PhiX174: 1
pBR322: 1
pUC57: 1
pUC18/19: 1
pTZ19R/U: 1
M13mp18/19: 2

Overdigestion Assay:
No detectable change in the specific fragmentation pattern is observed after 160-fold overdigestion with Eco88I (10U/ug lambda DNA x 16 hours).

Ligation/Recutting Assay:
The ligation and recleavage assay was replaced with L0 test after validating experiments showed L0 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 Eco88I for 4 hours.

Blue/White Cloning Assay:
The B/W assay was replaced with L0 test after validating experiments showed L0 test ability to detect nuclease and phosphatase activities with sensitivity that equals to that of B/W test.

Storage and Stability:
May be stored at 4°C. For long-term storage, aliquot and store at -20°C. Aliquots are stable for 6 months after receipt. For maximum recovery of product, centrifuge the original vial prior to removing the cap. Further dilutions can be made in assay buffer.