SbfI (Restriction Enzyme)

REF: EG15568S

Recognition & Cleavage

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

Cleavage pattern:

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

Type: II restriction endonuclease · Common use: Routine cloning and fast DNA digestion

Isoschizomers: SdaI, Sse8387I (methylation sensitivities may differ)

Specs

Optimal temperature: 37 °C
Fast digest: 5–15 min in 1× CutOne® / CutOne® Color Buffer
Heat inactivation: 80 °C · 20 min
Storage: −20 °C

Pack Components

Component	EG15568S
SbfI (20 U/µl)	25 µl (500 U)
10× CutOne® Buffer	1 ml
10× CutOne® Color Buffer	1 ml

Recommended Reaction

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

CutOne® Color Buffer includes density reagent and 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 SbfI.
30 µl (PCR product): 16 µl ddH₂O, 3 µl 10× CutOne®, ~0.2 µg DNA, 1 µl SbfI.
If PCR is unpurified, use 2 µl 10× buffer.
50 µl (genomic): 30 µl ddH₂O, 5 µl 10× CutOne®, 10 µl DNA (5 µg), 5 µl SbfI.
Mix gently, quick-spin; incubate at 37 °C for the indicated time.
(Optional) Heat-inactivate at 80 °C for 20 min or load gel directly (Color Buffer).

Notes

Total enzyme volume ≤10% of reaction to minimize glycerol-induced star activity.
Remove inhibitors (phenol, ethanol, EDTA, high salt, detergents) if digestion is incomplete.
Increase incubation time if total volume >20 µl.

Quality Control

Functional test: 1 µg λDNA + 1 µl SbfI in 20 µl CutOne® → complete digest in 15 min at 37 °C.
Prolonged incubation/star activity: 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 fragments recut.
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	0	pUC57	0
pUC18/19	1	SV40	0
M13mp18/19	1	Adeno2	3

Activity in Different Buffers

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

*Activities per the functional test conditions.

Write a review

Your Name

Your Review

Note: HTML is not translated!

Rating Bad Good

Captcha

Please complete the captcha validation below

SbfI

Brand: Best Enzymes
Product CAT#: EG15568S
Availability: In Stock

Available Options

Enzymes Handling Fee

Overnight Shipping Handling (+$29.99)

Size

Qty

0 reviews / Write a review