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SpeI, (SpeI)

Marka:EURx

Ürün Ölçü Fiyat TL Miktar
SpeI
E2398-01
500 u 65,6 EUR
+%20 KDV
3.227,52 TL
SpeI
E2398-02
2 500 u 259,2 EUR
+%20 KDV
12.752,64 TL

(Streptomyces phaeochromogenes)

5’-G C A T G C-3’
3’-C G T A C G-5’

Reaction Temperature: 37°C

Prototype: SphI

Inactivation Temperature (20 min): 65°C

Reaction Buffer:

1x Medium Buffer: 10 mM Tris-HCl (pH 7.5 at 37°C), 1 mM dithiothreitol, 10 mM MgCl2, 50 mM NaCl, 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 and 1 µg of lambda DNA. Incubation is at 37°C for 1 hr in a reaction volume of 50 µl.

Storage Buffer: 20 mM Tris-HCl (pH 7.5 at 22°C), 100 mM KCl, 0.1 mM EDTA, 5 mM β-mercaptoethanol, 100 µg/ml bovine serum albumin and 50% (v/v) glycerol.

Storage Conditions: Store at –20°C.

Quality Control:

Non-specyfic Endonucleases: Incubation of 40 units of SphI with 1 µg of lambda DNA at 37°C for 5 hrs (a 200-fold over-digestion) resulted in the same sharp characteristic banding pattern as the standard assay reaction, as determined by agarose gel electrophoresis.

Nicking Activity (phiX-174): Incubation of 20 units of SphI with 1 µg of phiX-174 DNA at 37°C for 5 hrs resulted in <5% conversion from Form I to II and <0% conversion from Form II to III as determined by agarose gel electrophoresis.

3’-Exonuclease: 5, 10 and 20 units of SphI 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 0.02-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 SphI with 0.05 µg 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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