BamH I | Enzimas "B" Cellco

BamH I

Ficha técnica

Código	BamH I
Categoria	Enzimas de Restrição
Marca	Cellco

Descrição Geral

BamH I | Enzimas "B" Cellco
JBSpeed Restriction Enzyme

For in vitro use only!

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.

Source:
Bacillus amyloliquefaciens H

Shipping:

shipped on blue ice

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

Supplied with:

10x Universal Buffer (UB)

Recommended 50 μl assay

Components	50 µL rxn
10X Universal Buffer (UB)	5 µL
pure DNA1 or PCR product2	1 μg
enzyme	10 units
PCR grade water	fill up to 50 μl

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.

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)

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. #DPK-106)

- Gel Electrophoresis and Single Band Excision (e.g. Agarose Gel Extraction Kit, Cat.#DPK-105)

- Phenol-Chloroform Extraction or Ethanol Precipitation.

Reaction Enzymes Buffer Guide:

Buffer 1	10 × B1	100 mM 100 mM 1000 μg/ml	Tris-HCl (pH 7.9, 25°C) MgCl2 BSA
Buffer 2	10 × B2	100 mM 100 mM 500 mM 1000 μg/ml	Tris-HCl (pH 7.9, 25°C) MgCl2 NaCl BSA
Buffer 3	10 × B3	500 mM 100 mM 1000 mM 1000 μg/ml	Tris-HCl (pH 7.9, 25°C) MgCl2 NaCl BSA
Buffer 4	10 × B4	100 mM 100 mM 1500 mM 1000 μg/ml	Tris-HCl (pH 7.9, 25°C) MgCl2 NaCl BSA
Buffer 5	10 × B5	200 mM 100 mM 500 mM 1000 μg/ml	Tris-acetate (pH 7.9, 25°C) Mg-acetate K-acetate BSA

Reaction Buffer Compatibility:

Our restriction enzymes are fully compatible to restrictases and buffer systems from other manufacturers and can be used along in double digestions. To obtain best results, consult the corresponding manuals of all involved products.

Ligation and recutting:

After 50-fold overdigestion with BamH I, >95% of the DNA fragments can be ligated and recut with this enzyme.

Star activity:
Conditions of low ionic strength, high enzyme concentration, glycerol concentration >5 % or pH >8.0 may result in star activity.

DNA Methylation:
No Inhibition: dam, dcm, CpG

Quality Control:
All preparations are assayed for contaminating endonuclease, 3’-exonuclease, 5’-exonuclease/ 5’-phosphatase, as well as nonspecific single- and doublestranded DNase activities.

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BamH I | Enzimas "B" Cellco

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