BamHI-7500 Units
Description
7500 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: shipped on gel packs
Storage Conditions: store at -20 °C
avoid freeze/thaw cycles
Shelf Life: 12 months
Form: liquid (Supplied in 10 mM Tris-HCl pH 7.4, 50 mM KCl, 0.1 mM EDTA, 1 mM DTT, 200 μg/ml BSA and 50 % [v/v] glycerol)
Concentration: 10 units/μl
Source: Bacillus amyloliquefaciens H
Supplied with: 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 |
1 Supercoiled or high molecular weight DNA (e.g. plant genomic DNA) may require longer incubation time or higher amount of enzyme.
2 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 80 °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 – 75 % Relative Activity
B2 – 75-100 % Relative Activity
B3 – 100 % Relative Activity
B4 – 50-75 % Relative Activity
B5 – 75 % Relative Activity
1x UB – 100 % Relative Activity (recommended)
Jena Bioscience
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