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SfiI,(Sfil)

Marka:EURx

Ürün Ölçü Fiyat TL Miktar
SfiI
E2380-01
2 000 u 65,6 EUR
+%20 KDV
3.227,52 TL
SfiI
E2380-02
10 000 u 259,2 EUR
+%20 KDV
12.752,64 TL

(Streptomyces fimbriatus)

5’-G G C C N N N N N G G C C-3’
3’-C C G G N N N N N C C G G-5’

Reaction Temperature: 50°C

Prototype: SfiI

Inactivation Temperature (20 min): no

Reaction Buffer:

1x Medium Buffer: 10 mM Tris-HCl (pH 7.5 at 37°C), 10 mM MgCl2, 50 mM NaCl, 1 mM dithiothreitol, 100 µg/ml bovine serum albumin.

Unit Definition: One unit is the amount of enzyme required to complete 98% digestion of 1 µg of Ad-2 DNA in 1 hr in a total reaction volume of 50 µl.

Assay Conditions: 10 mM Tris-HCl (pH 7.5 at 37°C), 50 mM NaCl, 10 mM MgCl2, 1 mM dithiothreitol, 1 µg of Ad-2 DNA and 100 µg/ml bovine serum albumin. Incubation is at 50°C for 1 hr in a reaction volume of 50 µl.

Storage Buffer: 10 mM Tris-HCl (pH 7.5 at 22°C), 50 mM KCl, 0.1 mM EDTA, 1 mM dithiothreitol, 200 µg/ml bovine serum albumin and 50% (v/v) glycerol.

Storage Conditions: Store at –20°C.

Quality Control:

Non-specific Endonuclease: Incubation of 10 units of SfiI with 1 µg of Ad-2 DNA at 50°C for 16 hrs (a 160-fold over-digestion) resulted in the same sharp characteristic banding pattern as the standard assay reaction, as determined by agarose gel electrophoresis.

3'-Exonuclease: 5, 10 and 20 units of SfiI and 0.09 µg (0.65 pmol of 3'-ends) of lambda/TaqI fragments (3'-labeled with T4 DNA Polymerase and [³H]dGTP and [³H]dCTP), incubated for 1 hr at 50°C resulted in 0.03 slope of %-end label released per unit of enzyme. Reaction volume 10 µl.

5'-Exonuclease/5'-Phosphatase: Incubation of 5, 10 and 20 units of SfiI with 0.08 µg (0.74 pmol of 5'-ends) of [5'-³²P] lambda/HaeIII fragments for 1 hr at 50°C resulted in a less than 0.04-slope of %-end label released per unit of enzyme. Reaction volume 10 µl.

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