ApaLI (Restriction Enzyme)

REF: EG15601S

Recognition & Cleavage

Recognition site: 5′-GTGCAC-3′ (palindromic)

Cleavage pattern:

5′ … G^▼T G C A C … 3′
3′ … C A C G T_▲G … 5′

Equivalent slash notation: 5′ … G/TGCAC … 3′ and 3′ … CACGT/G … 5′ (generates a 5′ four-base overhang: TGCA).

Type: II restriction endonuclease · Common use: routine cloning & restriction mapping.

Isoschizomers: Alw44I, VneI (methylation sensitivities may differ).

Specs

Optimal temperature: 37 °C
Fast digest: 5–15 min in 1× CutOne® / CutOne® Color Buffer
Heat inactivation: Yes — typically 65–80 °C for 20 min (check lot COA)
Storage: −20 °C

Pack Components

ApaLI Kit Components
Component	EG15601S
ApaLI (10 U/µl)	200 µl (2000 U)
10× CutOne® Buffer	2 × 1 ml
10× CutOne® Color Buffer	2 × 1 ml

Specs & components subject to change per lot COA.

Recommended Reaction

Buffer: 1× CutOne® or 1× CutOne® Color Buffer
Incubation: 37 °C
Guideline: Use the fast-digestion setup below; scale volumes as needed.

Color Buffer includes density reagent + tracking dyes for direct gel loading.

Quick Protocol (Fast DNA Digestion)

20 µl (plasmid): 15 µl ddH₂O, 2 µl 10× buffer, ≤1 µg DNA, 1 µl ApaLI.
30 µl (PCR product): 16 µl ddH₂O, 3 µl 10× buffer, ∼0.2 µg DNA, 1 µl ApaLI.
If PCR is unpurified, use 2 µl 10× buffer (residual metal ions).
50 µl (genomic DNA): 30 µl ddH₂O, 5 µl 10× buffer, 10 µl DNA (5 µg), 5 µl ApaLI.
Mix gently, quick-spin; incubate at 37 °C for 15 min (plasmid), 15–30 min (PCR), or 30–60 min (genomic).
(Optional) Heat-inactivate at 65–80 °C for 20 min; load directly if Color Buffer was used.

Double / Multiple Digests

Use ~1 µl of each enzyme; keep total enzyme volume ≤10% of reaction.
If temperatures differ, start at the lower temperature, then add the second enzyme and shift higher.
Increase incubation time if total volume > 20 µl.

Quality Control

Functional test: 1 µg λ DNA (contains multiple ApaLI sites) + 1 µl ApaLI in 20 µl buffer → complete digest in ≤30 min at 37 °C.
Prolonged incubation / star activity: 3 h at 37 °C shows no non-specific degradation under recommended conditions.
Ligation & recutting: After over-digest, >95% ligation at 22 °C; >95% of ligated fragments recut by ApaLI.
Non-specific endonuclease: <10% nicked/linearized plasmid after 4 h at 37 °C.

Methylation Effects

Methylation sensitivity
Methylase	Effect
Dam	No effect
Dcm	No effect
CpG	Blocked (overlapping CpG)
EcoKI	Impaired
EcoBI	No effect

See product datasheet for host strain–specific methylation details.

Recognition Sites in Common DNAs

Approximate ApaLI site counts
DNA	# Sites	DNA	# Sites
λ DNA	4	ΦX174	1
pBR322	3	pUC57	3
pUC18/19	3	SV40	0
M13mp18/19	1	Adeno2	7

Counts are typical references; confirm with your construct.

Activity in Different Buffers

Relative activity by buffer
Buffer	Activity
CutOne®	100%
Thermo FastDigest	75%
NEB rCutSmart™	100%
Takara QuickCut™	100%

Values are guide-level; verify for your DNA/substrate.

Write a review

Your Name

Your Review

Note: HTML is not translated!

Rating Bad Good

Captcha

Please complete the captcha validation below

ApaLI

Brand: Best Enzymes
Product CAT#: EG15601S
Availability: In Stock

Available Options

Enzymes Handling Fee

Overnight Shipping Handling (+$29.99)

Size

Qty

0 reviews / Write a review