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For general laboratory use.
Please centrifuge briefly before opening (volume ≤2 ml).Unit Definition: One unit is defined as the amount of enzyme required to convert 10 nmoles of dNTPs to an acid insoluble form in 30 minutes at 37 °C.
Shipping: shipped on gel packs
Storage Conditions: store at -20 °C
avoid freeze/thaw cyclesShelf Life: 12 months
Form: liquid (Supplied in 100 mM KPO4 pH 6.5, 1 mM DTT and 50 % [v/v] glycerol)
Concentration: 5 units/μl
Applications:
- Fill-in of 5' overhangs to form blunt ends
- Removal of 3' overhangs to form blunt ends
Description:
Klenow Fragment is the large fragment of DNA Polymerase I that retains its 5'→3' polymerase, 3'→5' exonuclease and strand displacement activities. The enzyme lacks the 5'→3' exonuclease activity of intact DNA polymerase I. Klenow retains the polymerization fidelity of the holoenzyme without degrading 5' termini.Reaction conditions:
- Dissolve 0.1 - 4 μg of digested DNA in 1x Reaction Buffer supplemented with 40 μM each dNTP
- Add 1 unit Klenow Fragment per μg DNA
Incubate for 15 min. at 25 °C
Stop reaction by alternativly
- add EDTA to 10 mM final concentration
- Heat inactivation: 20 min. at 75 °C
10x Reaction Buffer:
500 mM Tris-HCl pH 7.6 at 25 °C, 50 mM MgCl2 and 10 mM DTT.Note:
Excessive amounts of enzyme or longer reaction times may result in recessed ends due to the 3'→5' exonuclease activity of the enzyme.Quality Control:
The enzyme is greater than 98 % pure as indicated by SDS-polyacrylamide gel electrophoresis and contains no detected endonuclease activity. Incubation of 10 units of Klenow with supercoiled plasmid DNA produced no nicked molecules after 20 hours at 37 °C as determined by agarose gel electrophoresis analysis. -
产品组分
内容
型号
规格 储存温度
克列诺片段(Klenow Fragment)
EN148-01 300Units
-20°C 克列诺片段(Klenow Fragment)
EN148-02 1500Units
-20°C 操作手册
1 1 常温
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营销中心
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注意事项
保存建议 厂家推荐蓝冰运输。当您收到产品后，按照说明书建议保存于-20°C。 -
FAQ
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