FastDigest® PpuMI (Psp5II)

Kód produktu: FD0764 Kód výrobce: FD0764 Kód dodavatele: {947A1FB8-F54C-4A6A-93EA-EF23A2052A72} Výrobce: UAB Fermentas
2 250,60 Kč
1 860,00 Kč bez DPH
do týdne
50 rxns

5'...R GG W C C Y...3'
3'...Y C C W GG R...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.

Isoschizomers Search for commercial isoschizomers using REsearch.
Compatible Ends FastDigestAvaII (Eco47I), FastDigest RsrII (CpoI), FastDigest SanDI (KflI), FastDigest Sau96I (Cfr13I), AvaII, Cfr13I, CpoI, Eco47I, SanDI, Sau96I.
Formulation 1 μL of enzyme (1 FDU) cleaves 1 μg of lambda DNA/CpoI fragments in 5 minutes at 37°C in 1X FastDigest Buffer.
Hazardous No
Recommended Reaction Conditions

1X FastDigest buffer or 1X FastDigest Green buffer at 37°C.
1 µL of FastDigest PpuMI (Psp5II) is formulated to digest up to:

  • 1 µg of lambda DNA/CpoI fragments in 5 minutes
  • 1 µg of plasmid DNA in 5 minutes
  • 0.2 µg of PCR product in 5 minutes
  • 1 µg of genomic DNA in 20 minutes or 5 µg of genomic DNA in 2 hours
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 5 5 20 2 80°C, 5 min 6

Methylation Effects

  • Dam: never overlaps - no effect.
  • Dcm: may overlap - blocked.
  • CpG: never overlaps - no effect.
  • EcoKI: never overlaps - no effect.
  • EcoBI: may overlap - effect not determined.
Methylation type Sequence Cleavage effect
Dcm(CCWGG) 5'...RGGWCm5CT GG...3'
3'...YCCWG GAm5CC...5'
Blocked

Number of recognition sites in DNA molecules

Lambda ΦX174 M13mp18/19 pBR322 puc18/19 pUC57
3 0 0 2 0 0
pTZ19R/U pTZ57R pBluescriptIIKS(±) pBluescriptIISK(±) pACYC177 pACYC184
0 0 0 0 0 2

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
AG^GACCY
  • CpoI (RsrII)/FastDigest RsrII (CpoI)* (CG^GACCG)
  • Eco47I (AvaII)/FastDigest AvaII (Eco47I)* (G^GACC)
  • BmgT120I
  • Cfr13I (Sau96I)/FastDigest Sau96I (Cfr13I)
  • Eco47I (AvaII)/FastDigest AvaII (Eco47I)
  • FastDigest SanDI (KflI)* (GG^GACCC)
  • BmgT120I
  • BspLI (NlaIV)/FastDigest NlaIV (BspLI)
  • Cfr13I (Sau96I)/FastDigest Sau96I (Cfr13I)
  • Eco47I (AvaII)/FastDigest AvaII (Eco47I)
  • EcoO109I (DraII)/FastDigest EcoO109I
  • Psp5II (PpuMI)/FastDigest PpuMI (Psp5II)
AG^GTCCY
  • CpoI (RsrII)/FastDigest RsrII (CpoI)* (CG^GTCCG)
  • Eco47I (AvaII)/FastDigest AvaII (Eco47I)* (G^GTCC)
  • BmgT120I
  • Cfr13I (Sau96I)/FastDigest Sau96I (Cfr13I)
  • Eco47I (AvaII)/FastDigest AvaII (Eco47I)
  • SetI
  • FastDigest SanDI (KflI)* (GG^GTCCC)
  • BmgT120I
  • BspLI (NlaIV)/FastDigest NlaIV (BspLI)
  • Cfr13I (Sau96I)/FastDigest Sau96I (Cfr13I)
  • Eco47I (AvaII)/FastDigest AvaII (Eco47I)
  • EcoO109I (DraII)/FastDigest EcoO109I
  • Psp5II (PpuMI)/FastDigest PpuMI (Psp5II)
  • SetI
GG^GACCY
  • CpoI (RsrII)/FastDigest RsrII (CpoI)* (CG^GACCG)
  • Eco47I (AvaII)/FastDigest AvaII (Eco47I)* (G^GACC)
  • BmgT120I
  • BspLI (NlaIV)/FastDigest NlaIV (BspLI)
  • Cfr13I (Sau96I)/FastDigest Sau96I (Cfr13I)
  • Eco47I (AvaII)/FastDigest AvaII (Eco47I)
  • FaqI (BsmFI)/FastDigest BsmFI (FaqI)
  • FastDigest SanDI (KflI)* (GG^GACCC)
  • BmgT120I
  • BspLI (NlaIV)/FastDigest NlaIV (BspLI)
  • Cfr13I (Sau96I)/FastDigest Sau96I (Cfr13I)
  • Eco47I (AvaII)/FastDigest AvaII (Eco47I)
  • EcoO109I (DraII)/FastDigest EcoO109I
  • FaqI (BsmFI)/FastDigest BsmFI (FaqI)
  • FastDigest SanDI (KflI)
  • Psp5II (PpuMI)/FastDigest PpuMI (Psp5II)
GG^GTCCY
  • CpoI (RsrII)/FastDigest RsrII (CpoI)* (CG^GTCCG)
  • Eco47I (AvaII)/FastDigest AvaII (Eco47I)* (G^GTCC)
  • BmgT120I
  • BspLI (NlaIV)/FastDigest NlaIV (BspLI)
  • Cfr13I (Sau96I)/FastDigest Sau96I (Cfr13I)
  • Eco47I (AvaII)/FastDigest AvaII (Eco47I)
  • FastDigest SanDI (KflI)* (GG^GTCCC)
  • BmgT120I
  • BspLI (NlaIV)/FastDigest NlaIV (BspLI)
  • Cfr13I (Sau96I)/FastDigest Sau96I (Cfr13I)
  • Eco47I (AvaII)/FastDigest AvaII (Eco47I)
  • EcoO109I (DraII)/FastDigest EcoO109I
  • FastDigest SanDI (KflI)
  • Psp5II (PpuMI)/FastDigest PpuMI (Psp5II)

Obsah balení

50 rxns

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