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MspI, (Hapll)

Marka:EURx

Ürün Ölçü Fiyat TL Miktar
MspI
E2290-01
5 000 U 57,6 EUR
+%20 KDV
2.833,92 TL
MspI
E2290-02
25000U 228,8 EUR
+%20 KDV
11.256,96 TL

(Moraxella species)

5’-C C G G-3’
3’-G G C C-5’

Reaction Temperature: 37°C

Prototype: HpaII

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.

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, 100 µg/ml bovine serum albumin, 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 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-specific Endonuclease: Incubation of 20 units of MspI with 1 µg of 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 MspI and 0.010 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 0.04 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 MspI with 0.05 µg of 5'-ends of [5'-³³P]lambda/HaeIII fragments for 1 hr at 37°C resulted in - 0.06 slope of %-end label released per unit of enzyme. Reaction volume 10 µl.

 

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