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Key Documents

SPEI-RO

Roche

Spe I

from Sphaerotilus species

同義詞:

Spe I, SPE I

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About This Item

分類程式碼代碼:
12352204

生物源

bacterial (Sphaerotilus spp.)

品質等級

形狀

solution

比活性

10000 U/mL

包裝

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])

製造商/商標名

Roche

參數

37 °C optimum reaction temp.

顏色

colorless

pH值

8.0 (39 °F)

溶解度

water: miscible

適合性

suitable for molecular biology

應用

life science and biopharma
sample preparation

異物活動

Endonucleases 10 units, none detected

運輸包裝

dry ice

儲存溫度

−20°C

一般說明

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

特異性

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).

品質

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分析

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

單位定義

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.

儲存和穩定性

Do not store below −25°C

分析報告

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

其他說明

仅用于生命科学研究。不可用于诊断。

僅套裝組件

產品號碼
描述

  • Enzyme Solution

  • SuRE/Cut Buffer H 10x concentrated

儲存類別代碼

12 - Non Combustible Liquids

水污染物質分類(WGK)

WGK 1

閃點(°F)

does not flash

閃點(°C)

does not flash


分析證明 (COA)

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文章

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

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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