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ApaI

5'...G G G C CC...3'
3'...CC C G G G...5'
Available as a FastDigest enzyme for rapid DNA digestion

B buffer for 100% activity FastDigest enzyme available Genome qualified Optimal incubation at 37°C LO certified Cleavage blocked or impaired by overlapping CpG methylation Cleavage blocked or impaired by overlapping Dcm methylation Recombinant enzyme Thermal inactivation at 65°C in 20 min

The ApaI restriction enzyme recognizes GGGCC^C sites and cuts best at 37°C in B buffer (Isoschizomers: PspOMI)
  
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Lambda DNA digested with ApaI

Lambda DNA 0.7% agarose, 1 cleavage site

Thermo Scientific conventional restriction endonucleases are a large collection of high quality restriction enzymes, optimized to work in one of the buffers of the Five Buffer System. In addition, the universal Tango buffer is provided for convenience in double digestions. All of the enzymes exhibit 100% activity in the recommended buffer and reaction conditions. To ensure consistent performance, Thermo Scientific restriction enzyme reaction buffers contain premixed BSA, which enhances the stability of many enzymes and binds contaminants that may be present in DNA preparations.

Highlights

  • Superior quality – stringent quality control and industry leading manufacturing process
  • Convenient color-coded Five Buffer System
  • Includes universal Tango buffer for double-digestions
  • BSA premixed in reaction buffers
  • Wide selection of restriction endonuclease specificities

Applications

  • Molecular cloning
  • Restriction site mapping
  • Genotyping
  • Southern Blot
  • Restriction fragment length polymorphism (RFLP)
  • SNP

Note

Incubation at 30°C results in a 2-fold increase in activity. ApaI is inhibited by salt concentrations above 50 mM. ApaI cleavage is impaired by overlapping dcm methylation. To avoid dcm methylation, use a dam-, dcm- strain, such as GM2163.

For methylation sensitivity refer to product specifications.

  
Storage Condition-20 C
HazardousNo
Compatible EndsBseSI(GGGCC^C), Eco24I(GGGCC^C), SduI(GGGCC^C).
Conditions for 100% Activity

1X Buffer B:

  • 10 mM Tris-HCl (pH 7.5 at 37°C), 10 mM MgCl2, and 0.1 mg/mL BSA
  • Incubate at 37°C
Digestion of Agarose-embedded DNAMinimum 5 units of the enzyme are required for complete digestion of 1 µg of agarose-embedded lambda DNA in 16 hours.
Double DigestionPerform double digestion using DoubleDigest.
IsoschizomersSearch for commercial isoschizomers using REsearch.
Storage Buffer

ApaI is supplied in:

10 mM Tris-HCl (pH 7.4 at 25°C), 50 mM NaCl, 1 mM DTT, 1 mM EDTA, 0.2 mg/mL BSA and 50% (v/v) glycerol.

Reaction conditions

Recommended buffer for 100% activity Optimal temperature Enzyme activity in Thermo Scientific buffers, % Tango buffer for double digestion
B (blue)

1X
G (green)

1X
O (orange)

1X
R (red)

1X
Tango (yellow)

1X / 2X
B (Blue) 37°C 100 20-50 0-20 0-20 20-50 0-20 1X

Methylation Effects

Methylation type Sequence Cleavage effect
Dcm(CCWGG)

5'...GGGCCm5CW GG...3'

3'...CCCGG GWm5CC...5'

Impaired
CpG

5'...GGGCCm5C G...3'

3'...CCCGG Gm5C...5'

Impaired
CpG

5'...m5C GGGCCm5C G...3';

3'... Gm5CCCGG Gm5C...5'

Impaired

Cleavage efficiency close to the termini of PCR fragments

bp from the recognition site to fragment end
1 2 3 4 5
50-100

Number of recognition sites in DNA molecules

Lambda ΦX174 M13mp18/19 pBR322 puc18/19 pUC57
1 0 0 0 0 1
pTZ19R/U pTZ57R pBluescriptIIKS(±) pBluescriptIISK(±) pACYC177 pACYC184
0 1 1 1 0 0

New sites generated by ligation

Newly generated recognition sites resulting from ligation of protruding compatible DNA ends

Recognition Sequence Second Enzyme Enzymes recognizing newly generated recognition sequence
GGGCC^C
  • BseSI (Bme1580I)/FastDigest Bme1580I (BseSI)* (GGGCC^C)
  • Eco24I (BanII)* (GGGCC^C)
  • SduI (Bsp1286I)/FastDigest Bsp1286I (SduI)* (GGGCC^C)
  • ApaI/FastDigest ApaI
  • BmgT120I
  • BseSI (Bme1580I)/FastDigest Bme1580I (BseSI)
  • Bsp120I (PspOMI)/FastDigest Bsp120I
  • BspLI (NlaIV)/FastDigest NlaIV (BspLI)
  • BsuRI (HaeIII)/FastDigest HaeIII (BsuRI)
  • Cfr13I (Sau96I)/FastDigest Sau96I (Cfr13I)
  • CviJI
  • Eco24I (BanII)
  • SduI (Bsp1286I)/FastDigest Bsp1286I (SduI)