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

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)