BamHI (Restriction Enzyme)

REF: EG15510S

Recognition & Cleavage

Recognition site: 5'-GGATCC-3' (palindromic)

Cleavage pattern:

5' ... G^▼G A T C C ... 3'
3' ... C C T A G_▲G ... 5'

Type: II restriction endonuclease · Common use: Routine cloning, vector construction, fast digests

Isoschizomers: — (not listed; methylation sensitivities may differ)

Specs

Optimal temperature: 37 °C
Fast digest: 5–15 min in 1× CutOne® / CutOne® Color Buffer
Heat inactivation: Not heat-inactivatable — purify digest before downstream steps
Storage: −20 °C

Pack Components

Component	EG15510S
BamHI (20 U/µl)	500 µl (10,000 U)
10× CutOne® Buffer	3 × 1 ml
10× CutOne® Color Buffer	3 × 1 ml

Recommended Reaction

Buffer: 1× CutOne® or 1× CutOne® Color Buffer
Incubation: 37 °C
Typical duration: 5–15 min (plasmid/PCR); 30–60 min (genomic DNA)

CutOne® Color Buffer includes density reagent + red/yellow tracking dyes for direct gel loading.

Quick Protocol (Fast DNA Digestion)

20 µl (plasmid): 15 µl ddH₂O, 2 µl 10× CutOne®, ≤1 µg DNA, 1 µl BamHI.
30 µl (PCR product): 16 µl ddH₂O, 3 µl 10× CutOne®, ~0.2 µg DNA, 1 µl BamHI.
If unpurified PCR, use 2 µl 10× buffer.
50 µl (genomic): 30 µl ddH₂O, 5 µl 10× CutOne®, 5 µg DNA, 5 µl BamHI.
Mix gently, quick-spin; incubate at 37 °C for the indicated time.
Post-digest: Purify the reaction (phenol/chloroform or column) before ligation/transformation.

Notes

Total enzyme volume ≤10% of reaction to avoid glycerol-induced star activity.
Contaminants (phenol, ethanol, EDTA, high salt, detergents) inhibit digestion—purify DNA if needed.
Increase incubation time if total volume >20 µl.

Quality Control

Functional test: Complete λDNA digestion in 15 min at 37 °C (20 µl, 1 µg DNA, 1 µl enzyme).
Prolonged incubation: 3 h at 37 °C shows no nuclease degradation; longer may cause star activity.
Ligation & recutting: >95% ligation at 22 °C (T4 DNA Ligase); >95% of ligated products recut by BamHI.
Non-specific endonuclease: <10% nicked/linearized plasmid after 4 h at 37 °C.
Blue/White assay: <1% white colonies (DNA ends remain intact).

Methylation Effects

Methylase	Effect
Dam	No effect
Dcm	No effect
CpG	No effect
EcoKI	No effect
EcoBI	No effect

Recognition Sites in Common DNAs

DNA	# Sites	DNA	# Sites
λDNA	5	ΦX174	0
pBR322	1	pUC57	1
pUC18/19	1	SV40	1
M13mp18/19	1	Adeno2	3

Buffer Compatibility (Activity)

Buffer	Activity
CutOne®	100%
NEB rCutSmart™	100%
Thermo FastDigest	100%
Takara QuickCut™	100%

*Activities per functional test conditions.

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BamHI

Brand: Best Enzymes
Product CAT#: EG15510S
Availability: In Stock

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BamHI (Restriction Enzyme)

Recognition & Cleavage

Recommended Reaction

Quick Protocol (Fast DNA Digestion)

Notes

Quality Control

Methylation Effects

Recognition Sites in Common DNAs

Buffer Compatibility (Activity)

Write a review

BamHI

Available Options

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