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SspI, (SspI)

Marka:EURx

Ürün Ölçü Fiyat TL Miktar
SspI
E2402-01
1 000 u 40 EUR
+%20 KDV
1.968,00 TL
SspI
E2402-02
5 000 u 259,2 EUR
+%20 KDV
12.752,64 TL

(Sphaerotilus species)

5’-A A T A T T-3’
3’-T T A T A A-5’

Reaction Temperature: 37°C

Prototype: SspI

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, 0,025% Triton X-100, 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: 10 mM Tris-HCl (pH 7.5 at 37°C), 50 mM NaCl, 10 mM MgCl2, 1 mM dithiothreitol, 0,025% Triton X-100, 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 22°C), 50 mM KCl, 0.1 mM EDTA, 1 mM dithiothreitol, 100 µg/ml bovine serum albumin and 50% (v/v) glycerol.

Storage Conditions: Store at –20°C.

Quality Control:

Non-specyfic Endonucleases: Incubation of 10 units of SspI with 1 µg lambda DNA at 37°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 SspI and 0.21 µg (1.61 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 0.04-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 SspI with 0.04 µg (0.37 pmol of 5’-ends) of [5’-³²P] lambda/HaeIII fragments for 1 hr at 37°C resulted in 0.02-slope of %-end label released per unit of enzyme. Reaction volume 10 µl.

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