EcoRI, 15000 Units
Description
15000 units
For general laboratory use.
Unit Definition: One unit is the amount of enzyme required to completely digest 1 μg of Lambda DNA (5 sites) in 1 hour in a total reaction volume of 50 μl. Enzyme activity was determined in the recommended reaction buffer.
Shipping:Â shipped on gel packs
Storage Conditions: store at -20 °C
avoid freeze/thaw cycles
Shelf Life:Â 12 months
Form: liquid (Supplied in 5 mM KPO4 pH 7.4, 300 mM NaCl, 0.1 mM EDTA, 1 mM DTT, 200 μg/ml BSA, 0.15 % Triton X-100 and 50 % [v/v] glycerol)
Concentration: 10 units/μl
Source: Escherichia coli RY 13
Supplied with:Â 10x Universal Buffer (UB)
Recommended 50 μl assay
| 5 μl | 10x Universal Buffer (UB) |
| 1 μg | pure DNA1 or PCR product2 |
| 10 units | enzyme |
| fill up to 50 μl | PCR grade water |
1Â Supercoiled or high molecular weight DNA (e.g. plant genomic DNA) may require longer incubation time or higher amount of enzyme.
2Â Some enzymes may require additional DNA bases flanking the restriction site for complete digestion.
Protocol:
- The enzyme should not exceed 10 % of total reaction volume.
- Add enzyme as last component. Mix components well before adding enzyme. After enzyme addition, mix gently by pipetting. Do not vortex.
- Incubate 5 to 10 min. at 37 °C.
- Stop reaction by alternatively:
– Addition of 2.1 μl EDTA pH 8.0 [0.5 M], final 20 mM
– Heat Inactivation (20 min. at 65 °C)
– Spin Column DNA Purification (e.g. PCR Purification Kit, Cat.-No. PP-201S/L)
– Gel Electrophoresis and Single Band Excision (e.g. Agarose Gel Extraction Kit, Cat.-No. PP-202S/L)
– Phenol-Chloroform Extraction or Ethanol Precipitation.
Double Digestion – Buffer Compatibility:
B1 – 25-50 % Relative Activity
B2 – 50-75 % Relative Activity
B3 – 75 % Relative Activity
B4 – 50-75 % Relative Activity
B5 – 75 % Relative Activity
1x UB – 100 % Relative Activity (recommended)
Jena Bioscience
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