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BanII, (HgiJII)

Marka:EURx

Ürün Ölçü Fiyat TL Miktar
BanII, (HgiJII)
E2060-01
1 000 u 65,6 EUR
+%20 KDV
3.227,52 TL
BanII, (HgiJII)
E2060-02
5 000 u 259,2 EUR
+%20 KDV
12.752,64 TL

BanII

(Bacillus aneurinolyticus)

5’-G Pu G C Py C-3’
3’-C Py C G Pu G-5’

Reaction Temperature: 37°C

Prototype: HgiJII

Inactivation Temperature (20 min): 65°C

Reaction Buffer:

1x Acet Buffer: 20 mM Tris-acetate (pH 7.5 at 37°C), 10 mM magnesium acetate, 50 mM potassium acetate, 1 mM dithiothreitol, 100 µg/ml bovine serum albumin.

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: 20 mM Tris-acetate (pH 7.5 at 37°C), 10 mM magnesium acetate, 50 mM potassium acetate, 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 7.5 at 4°C), 50 mM NaCl, 0.1 mM EDTA, 1 mM dithiothreitol, 0.1% Triton™X-100, 500 µg/ml bovine serum albumin and 50% (v/v) glycerol.

Storage Conditions: Store at –20°C.

Quality Control:

Non-specific Endonuclease: Incubation of 20 units of BanII with 1 µg of lambda DNA at 37°C for 16 hrs (a 320-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 BanII and 0.25 µg 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 0.11-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 BanII with 0.05 µg of 5'-ends of [5'-³²P]lambda/HaeIII fragments for 1 hr at 37°C resulted in 0.04-slope of %-end label released per unit of enzyme. Reaction volume 10 µl.

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