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Source
: Bacillus globigii ATCC 49760
Compatible ends
(NNN…3')
AccB7 I, Ade I, Afi I, AlwN I, BsaX I, Bsc4 I, BseL I, BsiY I, Bsl I, BstAP I, BstMW I, Cai I, Dra III, Mwo I, PflB I, PflM I, Sfi I, Van91 I
Isoschizomers
: Tsp8E 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 Bgl 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 'D'[final conc.: 6 mM Tris-HCl(pH 7.9 @ 37℃), 6 mM MgCl2, 150 mM NaCl, 1 mM DTT], 1~2 ㎕ of DNA(0.5~1.0 ㎍/㎕), 1 ㎕ of Bgl I and 42~43 ㎕ of sterile water in 50 ㎕ reaction mixture. Incubate at 37℃.
Heat Inactivation
: 65℃ for 20 min.
Activity in BeamsBio™ Buffer System
|
Rxn Buffer |
A |
B |
C |
D |
|
Activity(%) |
10~50 |
10~50 |
75~100 |
75~100 |
|
Activity in a complete PCR Mix : 10~50%
Conditions: [10 mM Tris-HCl(pH 8.3 @ 25℃), 50 mM KCl, 1.5 mM MgCl2, 2 pmol of primers, 200 μM dNTP Mix, 2.5 U Taq DNA polymerase and 1 ㎍ of DNA(template or substrate) in 50 ㎕ of reaction volume] After 30 amplication cycles, add 5 units of Bgl I. And then incubate at 37℃ for one hour. Relative activity is determined by gel electrophoresis.
Storage Conditions
: 10 mM Tris-HCl (pH 7.5 @ 25℃), 200 mM NaCl, 0.5 mM EDTA, 1 mM DTT, 500 ㎍/㎖ BSA, and 50% glycerol. Store at -20℃.
No. of Cleavage Sites
|
λ
|
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
|
|
29
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20
|
2
|
0
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3
|
1
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1
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Quality control
Overdigestion: The same band pattern as a digestion in one unit of Bgl I for one hour was showing after incubation of 1 ㎍ of λ DNA with units of Bgl 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 Bgl I.
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References
Pirrotta, V. Nucleic Acids Res. 3: 1747-1760 (1976)/ Duncan, C.H., Wilson, G.A. and Young, F.E. J. Bacteriol. 134: 338 (1978)/
Lee, Y.H., Chirikjian, G. J. Biol. Chem. 254: 6838-6841 (1979)/ Bickle, T.A., Pirrotta, V., Imber, R. Methods Enzymol. 65: 132-138 (1980)/
Burger, K.J., Schinzel, R. Mol. Gen. Genet. 189: 269-274 (1983)
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