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Merck

SPEI-RO

Roche

Spe I

from Sphaerotilus species

Sinónimos:

Spe I, SPE I

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About This Item

Código UNSPSC:
12352204

origen biológico

bacterial (Sphaerotilus spp.)

Nivel de calidad

formulario

solution

actividad específica

10000 U/mL

envase

pkg of 1,000 U (11008951001 [10 U/μl])
pkg of 1,000 U (11207644001 [40 U/μl])
pkg of 200 U (11008943001 [10 U/μl])

fabricante / nombre comercial

Roche

Parámetros

37 °C optimum reaction temp.

color

colorless

pH

8.0 (39 °F)

solubilidad

water: miscible

idoneidad

suitable for molecular biology

aplicaciones

life science and biopharma
sample preparation

actividad extraña

Endonucleases 10 units, none detected

Condiciones de envío

dry ice

temp. de almacenamiento

−20°C

Categorías relacionadas

Descripción general

Spe I recognizes the sequence *A↓*CTAGT and generates fragments with 5′-cohesive termini.

Especificidad

Recognition sites: *A*CTAGT
*A*CTAGT
Restriction site: *A↓*CTAGT
*A↓*CTAGT
Heat inactivation: Spe I can be heat inactivated by incubation at 65 °C for 15 minutes (up to 100 U/μg DNA).

Calidad

Absence of nonspecific endonuclease activities
1μg Ad2 DNA is incubated for 16 hours in 50μl SuRE/Cut Buffer H with an excess of Spe 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 Spe 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.

Perfil del ADN

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

Definición de unidad

One unit is the enzyme activity that completely cleaves 1 μg Ad2 DNA in one hour at +37 °C in a total volume of 25 μl SuRE/Cut Buffer H.

Almacenamiento y estabilidad

Do not store below −25°C

Nota de análisis

Compatible ends
Spe I ends are compatible with ends generated by Bln I, Nhe I, and Xba I.

Isoschizomers
The enzyme is an isoschizomer of Bcu I and Ahl I.

Methylation sensitivity
As indicated by (*) on the recognition sequence above, Spe I is inhibited by the presence of N6-methyladenine and 5′-methylcytosine ( mA↓m CTAGT).

Incubation temperature
+37°C

PFGE tested
Spe 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 using 10U of enzyme/μg DNA and 4 hour incubation time.

Ligation and recutting assay
Spe I fragments obtained by complete digestion of 1μg Ad2 DNA are ligated with 1U T4 DNA Ligase in a volume of 10μl by incubation for 16 hours at +4°C in 66mM Tris-HCl, 5mM MgCl2, 5mM Dithiothreitol, 1mM ATP, pH 7.5 (at +20°C), resulting in >90% recovery of Ad2 DNA.
Subsequent re-cutting with Spe I yields >95% of the typical pattern of Ad2 × Spe I fragments.
SuRE/Cut Buffer System
The buffer in bold is recommended for optimal activity
  • A: 75-100%
  • B: 75-100%
  • H: 100%
  • L: 75-100%
  • M: 100%

Activity in PCR buffer: Not tested

Otras notas

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

Solo componentes del kit

Referencia del producto
Descripción

  • Enzyme Solution

  • SuRE/Cut Buffer H 10x concentrated

Código de clase de almacenamiento

12 - Non Combustible Liquids

Clase de riesgo para el agua (WGK)

WGK 1

Punto de inflamabilidad (°F)

does not flash

Punto de inflamabilidad (°C)

does not flash


Certificados de análisis (COA)

Busque Certificados de análisis (COA) introduciendo el número de lote del producto. Los números de lote se encuentran en la etiqueta del producto después de las palabras «Lot» o «Batch»

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Artículos

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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