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Source
: Escherichia coli RFL52
Compatible ends
(5'…GGCC)
Aco I, BseX3 I, Bsp120 I, BstZ I, CciN I, Cfr I, Eae I, Eag I, EclX I, Gdi II, Not I, PspOM I, Xma III
Isoschizomers
: BseX3 I, BstZ I, Eag I, EclX I
Methylation Sensitivity
: Not sensitive to dam and dcm methylation
: Blocked by overlapping CpG methylation
Unit definition :
One Unit is defined as the amount of Eco52 I required to completely digest 1 ㎍ of λ DNA in one hour at 37℃ in 50 ㎕ assay mixture.
Typical Reaction Conditions
: Add 5 ㎕ of 10X Reaction Buffer 'Eco52 I'[final conc.: 10 mM Tris-HCl (pH 8.9 @ 37℃), 3 mM MgCl2, 100 mM NaCl], 1~2 ㎕ of DNA(0.5~1.0 ㎍/㎕), 5 ㎕ of 10X BSA(final conc.: 100 ㎍/㎖), 1 ㎕ of Eco52 I and 37~38 ㎕ of sterile water in 50 ㎕ reaction mixture. Incubate at 37℃.
Heat Inactivation
: 65℃ for 20 min.
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Activity in BeamsBio™ Buffer System
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Rxn Buffer |
A |
B |
C |
D |
'Eco52 I' |
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Activity(%) |
<10 |
<10 |
<10 |
<10 |
75~100 |
Storage Conditions
: 10 mM Tris-HCl (pH 8.0), 100 mM KCl, 0.1 mM EDTA, 1 mM DTT, 200 ㎍/㎖ BSA, and 50% glycerol. Store at -20℃.
No. of Cleavage Sites
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λ
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Ad2
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pUC18
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φX174
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pBR322
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M13mp18
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SV40
|
|
2
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19
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0
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0
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1
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0
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0
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Quality control
Overdigestion: The same band pattern as a digestion in one unit of Eco52 I for one hour was showing after incubation of 1 ㎍ of λ DNA with units of Eco52 I for 16 hr in 50 ㎕ of reaction mixture at 37℃ as determined by agarose gel electrophoresis.
Ligation-Recut: After a 10-fold digestion for one hour, % of λ DNA fragments were ligated with T4 DNA ligase at 16℃ and % of the ligated fragments could be recut with Eco52 I.
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References
Janulaitis, A., Stakenas, P.S., Petrusyte, M.P., Bitinaite, J.B., Klimasauskas, S.I., Butkus, V.V. Mol. Biol. (Mosk) 18: 115-129 (1983)
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