Storage: ˆ’20°C

UNSPSC Code: 12352200

RIDADR: NONH for all modes of transport

Analysis Note:
SuRE/Cut Buffer System
The buffer in bold is recommended for optimal activity
€¢ A: 100%
€¢ B: 100%
€¢ H: 100%
€¢ L: 25-50%
€¢ M: 100%

Application:
Bgl II has been used for the digestion of the genomic DNA.

General description:
Bgl II recognizes the sequence A†€œG°AT*CT and generates fragments with 5€²-cohesive termini. Bgl II is not inhibited by dam-methylation that overlaps the site indicated (°) on the recognition sequence, but is sensitive to 5-methyl and 5-hydroxymethylcytosine at the other site indicated (*). Bgl II is not known to have isoschizomers. Bgl II cannot be heat inactivated by incubating it for 15 minutes at +65°C. The incubation temperature is +37° C.

Other Notes:
For life science research only. Not for use in diagnostic procedures.

Quality:
Absence of nonspecific endonuclease activities
1 μg ?DNA is incubated for 16 hours in 50 μl SuRE/Cut Buffer M with excessBgl II. The number of enzyme units which do not change the enzyme-specific pattern is stated in the certificate of analysis.

Absence of exonuclease activity
Approximately 5 μg [3H] labeled calf thymus DNA are incubated with 3 μl Bgl II for 4 hours at +37°C in a total volume of 100 μl 50 mM Tris-HCl, 10 mM MgCl2, 1 mM Dithioerythritol, pH approximately 7.5. Under these conditions, no release of radioactivity is detectable, as stated in the certificate of analysis.

Compatibility:
Bgl II generates ends that are compatible with fragments generated by BamH I, Bcl I, Nde II, Sau 3A and Xho II.

DNA Profile:
Number of cleavage sites on different DNAs
€¢ Î»: 6
€¢ Ï†X174: 0
€¢ Ad2: 11
€¢ M13mp7: 1
€¢ pBR322: 0
€¢ pBR328: 0
€¢ pUC18: 0
€¢ SV40: 0

Preparation Note:
Relative activity in complete PCR mix

Relative activity in PCR mix (Taq DNA Polymerase buffer) is 0%. The PCR mix contained ?DNA, primers, 10 mM Tris-HCl (pH 8.3, +20°C), 50 mM KCl, 1.5 mM MgCl2, 200 μM dNTPs, 2.5 U Taq DNA polymerase. The mix was subjected to 25 amplification cycles. Activity in buffer of Pwo SuperYield DNA Polymerase PCR Mix is 85%. When supplemented with GC-RICH Solution activity is increased to 100%.

Ligation and recutting assay
Bgl II fragments obtained by complete digestion of 1 μg ?DNA are ligated with 1 U T4 DNA Ligase in a volume of 10 μl by incubation for 16 hours at +4°C in 66 mM Tris-HCl, 5 mM MgCl2, 5 mM Dithiothreitol, 1 mM ATP, pH 7.5 (at +20°C) resulting in >80% recovery of 1 μg ?DNA × Bgl II fragments.
Subsequent re-cutting with Bgl II yields >95% of the typical pattern of ?DNA × Bgl II fragments.

Specificity:
Recognition sites: AG°AT*CT
AG°AT*CT
Restriction site: A†“G°AT*CT
A†“G°AT*CT
Heat inactivation: No inactivation of Bgl II after incubation at 65 °C for 15 minutes.