Sma I

Кат. номер

10656348001

10220566001

11047639001

Sma I recognizes the sequence *C°C*CGGG and generates fragments with blunt ends.
Specificity
Recognition sites: *C °C*CGGG
*C °C*CGGG
Restriction site: *C °C*CGGG
*C °C*CGGG
Heat inactivation: Sma I can be heat inactivated by incubation at 65 °C for 15 minutes (up to 100 U/µg DNA).
Sma I has been used in macrorestriction analysis. It has also been used in the restriction enzyme mixture during restriction digestion, amid rapid pulsed-field gel electrophoresis (PFGE).
Quality
Absence of nonspecific endonuclease activities
1µg DNA is incubated for 16 hours in 50µl SuRE/Cut Buffer A with an excess of Sma 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 Sma 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

• : 3
• X174: 0
• Ad2: 12
• M13mp7: 0
• pBR322: 0
• pBR328: 0
• pUC18: 1
• SV40: 0

Unit Definition
One unit is the enzyme activity that completely cleaves 1 µg DNA in one hour at +25 °C in a total volume of 25 µl (1x) SuRE/Cut buffer A.
Analysis Note
Compatible ends
Sma I generates ends that are compatible with any blunt end.

Isoschizomers
The enzyme is an isoschizomer to Cfr9 I, PspA I, Xma I, and XmaC I.
Methylation sensitivity
Sma I is not inhibited by 5-methylcytosine at the middle of the three C residues (°) in the recognition sequence. However, the activity is inhibited by 5-methylcytosine at the other Cs (*) or 4-methylcytosine in any position within the recognition sequence (*C°C*CGGG).
Incubation temperature
+25°C
PFGE tested
Sma I has been tested in Pulsed-Field Gel Electrophoresis (on bacterial chromosomes). For cleavage of genomic DNA (E. coli C 600) embedded in agarose for PFGE analysis, we recommend 10 U of enzyme/µg DNA and 4 hour incubation time.

Ligation and recutting assay
Sma I fragments obtained by complete digestion of 1µg DNA are ligated with 1U T4 DNA Ligase in a volume of 10µl by incubation for 16 hours at +25°C in 66mM Tris-HCl, 5mM MgCl2, 5mM Dithioerythritol, and 1mM ATP, pH 7.5 (at +20°C) resulting in >80% recovery of 1µg DNA fragments.
Subsequent re-cutting with Sma I yields >80% of the typical pattern of DNA x Sma I fragments.
SuRE/Cut Buffer System
The buffer in bold is recommended for optimal activity

• A: 100%
• B: 0-10%
• H: 0-10%
• L: 0-10%
• M: 0-10%

Activity in PCR buffer: 100%
Relative activity in PCR mix (Taq DNA Polymerase buffer) is 100%. 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 reaction buffer of Pwo SuperYield DNA Polymerase PCR Mix is 100%. If supplemented with GC-RICH Solution, activity remains at 100%. Pwo SuperYield DNA Polymerase PCR Mix is not available in U.S.
For life science research only. Not for use in diagnostic procedures.

Quality Level	100,
form	solution
packaging	pkg of 1,000 U (10220566001 [10 U/µl]), pkg of 5,000 U (10656348001 [10 U/µl]), pkg of 5,000 U (11047639001 [40 U/µl])
Торговая марка	Roche
parameter	25 °C optimum reaction temp.
shipped in	dry ice
storage temp.	20°C

RIDADR

NONH for all modes of transport