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Apa I

Kód produktu: ER1411 Kód výrobce: ER1411 Kód dodavatele: {034B0F74-58FB-44B9-8F69-BC0C9C9722CB} Výrobce: Life Technologies Czech Republic s.r.o.
2 168,32 Kč
1 792,00 Kč bez DPH
do týdne
3000 units

5'...G G G C C↓C...3'
3'...C↑C C G G G...5'

Zobraz detailní popis
  • Zeptejte se odborníka

Detailní popis

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.

Compatible Ends BseSI(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 DNA Minimum 5 units of the enzyme are required for complete digestion of 1 µg of agarose-embedded lambda DNA in 16 hours.
Double Digestion Perform double digestion using DoubleDigest.
Hazardous No
Isoschizomers Search 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.

Storage Condition -20 C

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

  • Dam: never overlaps - no effect
  • Dcm: may overlap - cleavage impaired
  • CpG: may overlap - cleavage impaired
  • EcoKI: never overlaps - no effect
  • EcoBI: never overlaps - no effect
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)

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