Storage: −20°C
UNSPSC Code: 12352200
Components: Enzyme Solution; SuRE/Cut Buffer M 10x concentrated
RIDADR: NONH for all modes of transport
Analysis Note:
SuRE/Cut Buffer System
The buffer in bold is recommended for optimal activity
• A: 25-50%
• B: 25-50%
• H: 25-50%
• L: 75-100%
• M: 100%
Application:
Because of its unusual recognition sequence, Sfi I is useful for analyzing and cloning large DNA fragments.
Note: The octameric recognition sequence of Sfi I rarely occurs in the genomes of many organisms, especially those that have low G+C content.
General description:
Sfi I recognizes the sequence GG°C*CNNNN↓NGG°C°C and generates fragments with 3′-cohesive termini.
Note: Sfi I sites are a subset of Bgl I sites. The cleavage position in the N5 region is identical.
Other Notes:
For life science research only. Not for use in diagnostic procedures.
Quality:
Absence of nonspecific endonuclease activities
1μg Ad2 DNA is incubated for 16 hours in 50μl SuRE/Cut Buffer M with an excess of Sfi I. 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 5μg [3H] labeled calf thymus DNA are incubated with 3μl Sfi I for 4 hours at +37°C in a total volume of 100μl 50mM Tris-HCl, 10mM MgCl2, 1mM Dithioerythritol, pH approximately 7.5. Under these conditions, no release of radioactivity is detectable, as stated in the certificate of analysis.
DNA Profile:
Number of cleavage sites on different DNAs
• λ: 0
• φX174: 0
• Ad2: 3
• M13mp7: 0
• pBR322: 0
• pBR328: 0
• pUC18: 0
• SV40: 1
Specificity:
Recognition sites: GG °C*CN5GG °C °C
GG °C*CN5GG °C °CGG °C*CNNNNNGG °C °C
Restriction site: GG °C*CN4↓NGG °C °C
GG °C*CN4↓NGG °C °CGG °C*CNNNN↓NGG °C °C
Heat inactivation: No inactivation of Sfi I after incubation at 65 °C for 15 minutes.
Compatible ends
For an Sfi I end to be compatible and ligatable with any other end, the two corresponding recognition sites must have contained the same three internal Ns.
Isoschizomers
The enzyme has no known isoschizomers.
Methylation sensitivity
Sfi I is inhibited by the presence of 5-methylcytosine at the most central C residue (*) on the recognition sequence. 5-methylcytosine at the other C-positions (°) will not inhibit the enzyme.
Incubation temperature
+50°C