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SMAI-RO

Roche

Sma I

from Serratia marcescens Sb

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About This Item

UNSPSC-Code:
12352204

Biologische Quelle

Serratia marcescens

Qualitätsniveau

Form

solution

Spezifische Aktivität

10000 U/mL

Verpackung

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])

Hersteller/Markenname

Roche

Konzentration

<0.1 % (w/w)

Parameter

25 °C optimum reaction temp.

Methode(n)

electrophoresis: suitable

Farbe

colorless

pH-Wert

7.0 (39 °F)

Löslichkeit

water: miscible

Eignung

suitable for molecular biology

Anwendung(en)

life science and biopharma

Fremdaktivität

Endonucleases, none detected (up to 20 U with lambda-DNA)
Endonucleases, none detected (up to 20U with pBR 322-DNA )

Versandbedingung

dry ice

Lagertemp.

−20°C

Allgemeine Beschreibung

Sma I recognizes the sequence *C°C*C↓GGG and generates fragments with blunt ends.

Spezifität

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).

Anwendung

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).

Qualität

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.

DNA-Profil

Number of cleavage sites on different DNAs
  • λ: 3
  • φX174: 0
  • Ad2: 12
  • M13mp7: 0
  • pBR322: 0
  • pBR328: 0
  • pUC18: 1
  • SV40: 0

Einheitendefinition

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.

Lagerung und Haltbarkeit

Do not store below −25°C

Hinweis zur Analyse

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.
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 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.

Sonstige Hinweise

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

Nur Kit-Komponenten

Produkt-Nr.
Beschreibung

  • Enzyme Solution

  • SuRE/Cut Buffer A 10x concentrated

Lagerklassenschlüssel

12 - Non Combustible Liquids

WGK

WGK 1

Flammpunkt (°F)

does not flash

Flammpunkt (°C)

does not flash


Analysenzertifikate (COA)

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Die Dokumentenbibliothek aufrufen

Corinna Kehrenberg et al.
Antimicrobial agents and chemotherapy, 51(2), 483-487 (2006-12-06)
During a study of florfenicol-resistant porcine staphylococci from Denmark, the genes cfr and fexA were detected in the chromosomal DNA or on plasmids of Staphylococcus hyicus, Staphylococcus warneri, and Staphylococcus simulans. A novel variant of the phenicol resistance transposon Tn558
E M Ribot et al.
Journal of clinical microbiology, 39(5), 1889-1894 (2001-04-28)
We developed a rapid pulsed-field gel electrophoresis (PFGE) protocol for subtyping Campylobacter isolates based on the standardized protocols used by PulseNet laboratories for the subtyping of other food-borne bacterial pathogens. Various combinations of buffers, reagents, reaction conditions (e.g., cell suspension

Artikel

The term “Restriction enzyme” originated from the studies of Enterobacteria phage λ (lambda phage) in the laboratories of Werner Arber and Matthew Meselson.

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