FastDigest® AluI

Kód produktu: FD0014 Kód výrobce: FD0014 Kód dodavatele: {AE854437-B8A7-44F3-B140-5F45E81D6E4D} Výrobce: UAB Fermentas
2 260,28 Kč
1 868,00 Kč bez DPH
do týdne
100 react

.

5'...A GC T...3'
3'...T CG A...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.

No
Isoschizomers Search for commercial isoschizomers using REsearch.
Formulation 1 μL of enzyme (1 FDU) cleaves 1 μg of lambda DNA in 15 minutes at 37°C in 1X FastDigest Buffer.
Hazardous
Recommended Reaction Conditions

1X FastDigest buffer or 1X FastDigest Green buffer at 37°C.
1 µL of FastDigest AluI is formulated to digest up to:

  • 1 µg of lambda DNA in 15 minutes
  • 1 µg of plasmid DNA in 15 minutes
  • 0.2 µg of PCR product in 15 minutes
  • 1 µg of genomic DNA in 15 minutes or 5 µg of genomic DNA in 60 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 15 15 15 15 4 65°C, 5 min 16

Methylation Effects

    Dam: never overlaps - no effect.
    Dcm: never overlaps - no effect.
    CpG: never overlaps - no effect.
    EcoKI: may overlap - no effect.
    EcoBI: may overlap - blocked.
Methylation type Sequence Cleavage effect

EcoBI (TGA(N)8TGCT)

5'...TGm6AGCT(N)5 TGCT...3'
3'...AC TCGA(N)5m6ACGA...5'
Blocked

Number of recognition sites in DNA molecules

Lambda ΦX174 M13mp18/19 pBR322 puc18/19 pUC57
143 24 27 17 16 16
pTZ19R/U pTZ57R pBluescriptIIKS(±) pBluescriptIISK(±) pACYC177 pACYC184
17 17 17 17 12 13

New sites generated by ligation

Newly generated recognition sites resulting from ligation of blunt DNA ends

Recognition sequence Second restriction enzyme Restriction enzymes cleaving the newly generated recognition sequence
AG^CT
  • AjiI (BmgBI)* (CAC^GTC)
  • AjiI (BmgBI)* (GAC^GTG)
  • Eco105I (SnaBI)/FastDigest SnaBI (Eco105I) (TAC^GTA)
  • Eco72I (PmlI)/FastDigest PmlI (Eco72I) (CAC^GTG)
  • Ppu21I (BsaAI)/FastDigest BsaAI (Ppu21I)* (YAC^GTA)
  • Ppu21I (BsaAI)/FastDigest BsaAI (Ppu21I)* (YAC^GTG)
  • ZraI (GAC^GTC)
  • SetI
  • Bst1107I (BstZ17I)/FastDigest BstZ17I (Bst1107I) (GTA^TAC)
  • Csp6I (CviQI)/FastDigest Csp6I
  • RsaI/FastDigest RsaI
  • BsuRI (HaeIII)/FastDigest HaeIII (BsuRI) (GG^CC)
  • CviJI* (RG^CC)
  • Eco147I (StuI)/FastDigest StuI (Eco147I) (AGG^CCT)
  • Eco47III (AfeI)/FastDigest AfeI (Eco47III) (AGC^GCT)
  • MlsI (MscI)/FastDigest MscI (MlsI) (TGG^CCA)
  • CviJI
  • CviJI* (RG^CT)

Obsah balení

100 rxns

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