• Your shopping cart is empty!

  • SmaI

SmaI (Restriction Enzyme)

REF: EG15572S

Recognition & Cleavage

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

5′ ... C C C▼G G G ... 3′
3′ ... G G G▲C C C ... 5′

Type: II restriction endonuclease · Common use: cloning, mapping, and blunt-end ligation

Isoschizomers: Cfr9I, TspMI, XmaI (methylation sensitivities may differ)

Recommended Reaction

  • Buffer: 1× CutOne® or Color Buffer
  • Incubation: 25 °C (prolong incubation slightly if at 37 °C)
  • Durations: 15 min (plasmid DNA), 15–30 min (PCR), 30–60 min (genomic DNA)

CutOne® Color Buffer contains density reagent with red/yellow dyes for direct gel loading.

Quick Protocol (Fast DNA Digestion)

  1. 20 µl plasmid DNA: 15 µl H₂O, 2 µl 10× buffer, ≤1 µg DNA, 1 µl SmaI
  2. 30 µl PCR product: 16 µl H₂O, 3 µl 10× buffer, ~0.2 µg DNA, 1 µl SmaI
    If unpurified PCR, reduce buffer to 2 µl
  3. 50 µl genomic DNA: 30 µl H₂O, 5 µl 10× buffer, 5 µg DNA, 5 µl SmaI
  4. Incubate at 25 °C for indicated times
  5. (Optional) Heat-inactivate at 80 °C for 20 min; direct load possible if Color Buffer used

Double / Multiple Digests

  • Use ~1 µl each enzyme; total enzyme ≤10% of volume
  • If enzymes need different temps, start lower then shift higher
  • Increase incubation if total reaction >20 µl

Quality Control

  • Functional test: Complete λDNA digest in 15 min at 25 °C
  • Star activity: None detected in 3 h; longer incubation may show star activity
  • Ligation & recutting: >95% ligation with T4 ligase; >95% recutting
  • Non-specific nuclease: <10% nicked/linearized plasmid after 4 h
  • Blue/White assay: <1% white colonies, confirming end integrity

Methylation Effects

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

Recognition Sites in Common DNAs

DNA # Sites DNA # Sites
λDNA 3 ΦX174 0
pBR322 0 pUC57 1
pUC18/19 1 SV40 0
M13mp18/19 1 Adeno2 12

Activity in Different Buffers

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

Write a review

Note: HTML is not translated!
    Bad           Good
Captcha

SmaI

  • Brand: Best Enzymes
  • Product CAT#: EG15572S
  • Availability: In Stock

Available Options


0 reviews / Write a review

Related Products

DpnI

DpnI

DpnI (Type IIM Restriction Enzyme) REF: EG15585S Recognition & Cleavage Recogniti..

$0.00

BspQI

BspQI

BspQI (Type IIS Restriction Enzyme) REF: EG23503S Recognition & Cleavage Recognit..

$0.00

BsmBI

BsmBI

BsmBI (Type IIS Restriction Enzyme) REF: EG22507V Recognition & Cleavage Recognit..

$0.00

XhoI

XhoI

XhoI (Restriction Enzyme) REF: EG15583S Recognition & Cleavage Recognition site: ..

$0.00

SfiI

SfiI

SfiI (Restriction Enzyme) REF: EG15570S Recognition & Cleavage Recognition site: ..

$0.00

SbfI

SbfI

SbfI (Restriction Enzyme) REF: EG15568S Recognition & Cleavage Recognition site: ..

$0.00

SalI

SalI

SalI (Restriction Enzyme) REF: EG15566S Recognition & Cleavage Recognition site: ..

$0.00

SacI

SacI

SacI (Restriction Enzyme) REF: EG15565S Recognition & Cleavage Recognition site: ..

$0.00

NcoI

NcoI

NcoI (Restriction Enzyme) REF: EG15550S Recognition & Cleavage Recognition site: ..

$0.00

MluI

MluI

MluI (Restriction Enzyme) REF: EG15547S Recognition & Cleavage Recognition site: ..

$0.00

DpnII

DpnII

DpnII (Restriction Enzyme) REF: EG15533S Recognition & Cleavage Recognition site:..

$0.00

NsiI

NsiI

NsiI (Restriction Enzyme) REF: EG15606S Recognition & Cleavage Recognition site: ..

$0.00

PvuII

PvuII

PvuII (Restriction Enzyme) REF: EG15604S Recognition & Cleavage Recognition site:..

$0.00

SacII

SacII

SacII (Restriction Enzyme) REF: EG15605S Recognition & Cleavage Recognition site:..

$0.00

PacI

PacI

PacI (Restriction Enzyme) REF: EG15603S Recognition & Cleavage Recognition site: ..

$0.00

ApaLI

ApaLI

ApaLI (Restriction Enzyme) REF: EG15601S Recognition & Cleavage Recognition site:..

$0.00

BgIII

BgIII

BglII (Restriction Enzyme) REF: EG15591S Recognition & Cleavage Recognition site:..

$0.00

Tags: Restriction Enzyme