Sma I
ROCHE/SMAI-RO - from Serratia marcescens Sb
Product Type: Chemical
| application(s) | life science and biopharma |
| biological source | Serratia marcescens |
| color | colorless |
| concentration | <0.1 % (w/w) |
| foreign activity | Endonucleases, none detected (up to 20 U with lambda-DNA) |
| Endonucleases, none detected (up to 20U with pBR 322-DNA ) | |
| form | solution |
| manufacturer/tradename | Roche |
| 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]) | |
| parameter | 25 °C optimum reaction temp. |
| pH | 7.0 (39 °F) |
| Quality Level | 100  |
| shipped in | dry ice |
| solubility | water: miscible |
| specific activity | 10000 U/mL |
| storage temp. | −20°C |
| suitability | suitable for molecular biology |
| technique(s) | electrophoresis: suitable |
| 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*C↓GGG). 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 λDNA 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 λDNA fragments. Subsequent re-cutting with Sma I yields >80% of the typical pattern of λDNA × Sma I fragments. |
| Analysis Note: | 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% |
| Analysis Note: | Activity in PCR buffer: 100% Relative activity in PCR mix (Taq DNA Polymerase buffer) is 100%. The PCR mix contained λDNA, 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. |
| Application: | 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). |
| DNA Profile: | Number of cleavage sites on different DNAs • λ: 3 • φX174: 0 • Ad2: 12 • M13mp7: 0 • pBR322: 0 • pBR328: 0 • pUC18: 1 • SV40: 0 |
| General description: | Sma I recognizes the sequence *C°C*C↓GGG and generates fragments with blunt ends. |
| Other Notes: | For life science research only. Not for use in diagnostic procedures. |
| Quality: | Absence of nonspecific endonuclease activities 1μg λDNA 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. |
| Specificity: | Recognition sites: *C °C*CGGG *C °C*CGGG Restriction site: *C °C*C↓GGG *C °C*C↓GGG Heat inactivation: Sma I can be heat inactivated by incubation at 65 °C for 15 minutes (up to 100 U/μg DNA). |
| Storage and Stability: | Do not store below −25°C |
| Unit Definition: | One unit is the enzyme activity that completely cleaves 1 μg λDNA in one hour at +25 °C in a total volume of 25 μl (1x) SuRE/Cut buffer A. |
| RIDADR | NONH for all modes of transport |
| WGK Germany | WGK 1 |
| Flash Point(F) | does not flash |
| Flash Point(C) | does not flash |
| activity | specific activity: 10000 U/mL |
| Storage Temp. | −20°C |
| UNSPSC | 12352204 |
| Components | Enzyme Solution; SuRE/Cut Buffer A 10x concentrated |