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Potvrzuji
  1. Úvodní strana
  2. Reagencie
  3. Life Technologies / Fermentas
  4. FastDigest & CONVENTIONAL RESTRICTION ENZYMES
  5. FastDigest restrikční enzymy
  6. FastDigest® AleI (OliI)

FastDigest® AleI (OliI)

Kód produktu: FD1634 Kód výrobce: FD1634 Kód dodavatele: {8A86A739-2F2D-46D6-8644-66725554976E} Výrobce: Life Technologies Czech Republic s.r.o.
FastDigest® AleI (OliI)
2 480,50 Kč
2 050,00 Kč bez DPH
do týdne
20 rxns

5'...C A C N N↓N N G T G...3'
3'...G T G N N↑N N C A C...5'

Zobraz detailní popis

Detailní popis

Thermo Scientific FastDigest enzymes are an advanced line of fast restriction enzymes that are all 100% active in the universal FastDigest and FastDigest Green reaction buffers.

The universal buffer allows rapid single-, double- or multiple DNA digestion within 5-15 minutes eliminating any need for buffer change or subsequent DNA clean-up steps. DNA modifying enzymes, such as Klenow Fragment, T4 DNA Ligase, alkaline phosphatases and T4 DNA Polymerase all have 100% activity in FastDigest Buffer. Therefore, enzymes for downstream applications can be directly added to the FastDigest reaction mix.

For additional convenience FastDigest Green Buffer includes a density reagent and two tracking dyes for direct loading of digestion reaction products on gels.

Short incubation times and optimal composition of the universal FastDigest Buffer eliminate star activity effects.

Features

  • 100% activity of all FastDigest enzymes in the universal buffer
  • 100% buffer compatibility with downstream applications
  • Complete digestion in 5-15 minutes
  • Direct loading on gels
  • No star activity
  • 176 FastDigest enzymes available

Applications

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

Note

The FastDigest Green Buffer offers the same high performance in DNA digestion and downstream applications as the colorless FastDigest Buffer. For applications that require product analysis by fluorescence excitation (e.g. concentration measurements in UV light) the colorless FastDigest Buffer is recommended.

For methylation sensitivity refer to product specifications.

Formulation 1 μL of enzyme (1 FDU) cleaves 1 μg of ΦX174 DNA in 5 min at 37°C in 1X FastDigest Buffer.
Hazardous No
Isoschizomers Search for commercial isoschizomers using REsearch.
Recommended Reaction Conditions

1X FastDigest buffer or 1X FastDigest Green buffer at 37°C.
1 µL of FastDigest AleI (OliI) is formulated to digest up to:

  • 1 µg of phiX174 DNA in 5 minutes
  • 1 µg of plasmid DNA in 5 minutes
  • 0.2 µg of PCR product in 10 minutes
  • 1 µg of genomic DNA in 5 minutes or 5 µg of genomic DNA in 30 minutes
Storage Condition -20 C

Reaction conditions

Reaction temperature Digestion time with 1 µL of FastDigest enzyme, min bp from end of DNA required for complete digestion Thermal inactivation Incubation time without star activity, hours
Lambda, 1 µg/20 µL Plasmid DNA, 1 µg/20 µL PCR product, ~0.2 µg/30 µL Genomic DNA, 1 µg/10 µL
37°C 5
(phiX174 DNA)		 5 10 5 3 65°C, 5 min 6

Methylation Effects

  • Dam: never overlaps - no effect.
  • Dcm: never overlaps - no effect.
  • CpG: may overlap - cleavage impaired.
  • EcoKI: may overlap - blocked.
  • EcoBI: may overlap - blocked.
Methylation type Sequence Cleavage effect
CpG 5'...CAm5C GNNm5C GTG...3'
3'...GT Gm5CNN Gm5CAC...5'		 Impaired
CpG 5'...CAm5C GNNNGTG...3'
3'...GT Gm5CNNNCAC...5'		 No effect
EcoBI(TGA(N)8TGCT) 5'...TGm6A(N)4CACG TGCT...3'
3'...AC T(N)4GTGCm6ACGA...5'		 Blocked
EcoKI(AAC(N)6GTGC) 5'...Am6ACAC(N)4G TGC...3'
3'...T TGTG(N)4Cm6ACG...5'		 Blocked

Number of recognition sites in DNA molecules

Lambda ΦX174 M13mp18/19 pBR322 puc18/19 pUC57
20 1 0 0 0 0
pTZ19R/U pTZ57R pBluescriptIIKS(±) pBluescriptIISK(±) pACYC177 pACYC184
0 0 1 1 0 0

Obsah balení

20 rxns

Hodnocení produktu

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