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FokI, (FokI)

Marka:EURx

Ürün Ölçü Fiyat TL Miktar
FokI, (FokI)
E2170-01
500 u 57,6 EUR
+%20 KDV
2.833,92 TL
FokI, (FokI)
E2170-02
2 500 u 228,8 EUR
+%20 KDV
11.256,96 TL

FokI

(Flavobacterium okeanokoites)

5'-G G A T G (N)9-3'
   3’-C C T A C (N)13–5’

Reaction Temperature: 37°C

Prototype: FokI

Inactivation Temperature (20 min): 65°C

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.

It is not recommended to use more than 1 unit per 1 μg of DNA and incubate longer than 2 hours.

Unit Definition: One unit is the amount of enzyme required to completely digest 1 µg of lambda DNA in 1 hr in a total reaction volume of 50 μl.

Assay Conditions: 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 and 1 μg of lambda DNA. Incubation is at 37°C for 1 hr in a reaction volume of 50 µl.

Storage Buffer: 10 mM Tris-HCl (pH 7.4 at 25°C), 50 mM NaCl, 0.1 mM EDTA, 0.1 % Tween 20, 1 mM dithiothreitol, 200 µg/ml bovine serum albumin and 50% (v/v) glycerol.

Storage Conditions: Store at –20°C.

Quality Control:

Non-specific Endonucleases: Incubation of 10 units of FokI with 1 µg of lambda DNA at 37°C for 16 hrs resulted in the same sharp characteristic banding pattern as the standard assay reaction, as determined by agarose gel electrophoresis.

3’-Exonuclease: 2, 5 and 10 units of FokI and 0.16 µg (1.23 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 37°C resulted in 0.05-slope of %-end label released per unit of enzyme. Reaction volume 10 µl.

5’-Exonuclease/5’-Phosphatase: Incubation of 2, 5 and 10 units of FokI with 0.03 µg,(0.24 pmol of 5’-ends) of [5’-³²P] lambda/HaeIII fragments for 1 hr at 37°C resulted in 0.03-slope of %-end label released per unit of enzyme. Reaction volume 10 µl.

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