FastDigest® ApaLI (Alw44I)

Kód produktu: FD0044 Kód výrobce: FD0044 Kód dodavatele: {244FFF3B-B92B-42F7-A855-1BCED1B84FCF} Výrobce: UAB Fermentas
1 713,36 Kč
1 416,00 Kč bez DPH
do týdne
200 react

5'...GT G C A C...3'
3'...C A C G TG...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.

Search for commercial isoschizomers using REsearch.
Recommended Reaction Conditions
1X FastDigest buffer or 1X FastDigest Green buffer at
Compatible Ends FastDigest BfmI, BfmI.
Formulation 1 μL of enzyme (1 FDU) cleaves 1 μg of lambda DNA/SmaI fragments in 5 minutes at 37°C in 1X FastDigest Buffer.
Hazardous No
Isoschizomers

37°C.
1 µL of FastDigest ApaLI (Alw44II) is formulated to digest up to:

  • 1 µg of lambda DNA/SmaI fragments in 5 minutes
  • 1 µg of plasmid DNA in 60 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 60 10 5 4 80°C, 5 min 16

Methylation Effects

  • Dam: never overlaps - no effect.
  • Dcm: never overlaps - no effect.
  •    CpG: may overlap - blocked.
        EcoKI: may overlap - no effect.
        EcoBI: never overlaps - no effect.

        CpG 5'...GTGCAm5C G...3'
        3'...CACGT Gm5C...5' Blocked
        EcoKI(AAC(N)6GTGC) 5'...Am6AC(N)6G TGCAC...3'
        3'...T TG(N)6Cm6ACGTG...5' No effect

Methylation type Sequence Cleavage effect

Number of recognition sites in DNA molecules

Lambda ΦX174 M13mp18/19 pBR322 puc18/19 pUC57
4 1 1 3 3 3
pTZ19R/U pTZ57R pBluescriptIIKS(±) pBluescriptIISK(±) pACYC177 pACYC184
2 2 2 2 1 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
G^TGCAC
  • BfmI (SfcI)/FastDigest SfcI (BfmI)* (C^TGCAG)
  • BsgI
  • HpyCH4V

Newly generated recognition sites resulting from fill-in of 5'-overhang and self-ligation

Recognition Sequence Newly generated sequence after reaction Enzymes that cleave the newly generated sequence
G^TGCAC GTGCATGCAC
  • Cac8I
  • CviAII
  • FaiI
  • FatI
  • Hin1II (NlaIII)/FastDigest Hin1II
  • [HpyCH4V]
  • PaeI (SphI)/FastDigest PaeI
  • XceI (NspI)/FastDigest XceI

Obsah balení

200 rxns

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