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SPEI-RO

Roche

Spe I

from Sphaerotilus species

Synonym(s):

Spe I, SPE I

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About This Item

UNSPSC Code:
12352204

biological source

bacterial (Sphaerotilus spp.)

Quality Level

form

solution

specific activity

10000 U/mL

packaging

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])

manufacturer/tradename

Roche

parameter

37 °C optimum reaction temp.

color

colorless

pH

8.0 (39 °F)

solubility

water: miscible

suitability

suitable for molecular biology

application(s)

life science and biopharma
sample preparation

foreign activity

Endonucleases 10 units, none detected

shipped in

dry ice

storage temp.

−20°C

Related Categories

General description

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

Specificity

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).

Quality

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.

DNA Profile

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

Unit Definition

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.

Storage and Stability

Do not store below −25°C

Analysis Note

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

Other Notes

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

Kit Components Only

Product No.
Description

  • Enzyme Solution

  • SuRE/Cut Buffer H 10x concentrated

Storage Class Code

12 - Non Combustible Liquids

WGK

WGK 1

Flash Point(F)

does not flash

Flash Point(C)

does not flash


Certificates of Analysis (COA)

Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.

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Articles

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

Related Content

Restriction endonucleases in prokaryotes function primarily to protect against foreign genetic material, notably bacteriophage DNA.

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