FastDigest® PfoI

Kód produktu: FD1754 Kód výrobce: FD1754 Kód dodavatele: {9361DDC5-B5E5-4956-929D-2489D28D9DDB} Výrobce: UAB Fermentas
5 027,55 Kč
4 155,00 Kč bez DPH
do týdne
20 rxns

5'...TC C N G G A...3'
3'...A G G N C CT...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.

Compatible Ends FastDigest SexAI (CsiI), EcoRII, SexAI, BssKI.
Formulation 1 µL of enzyme (1 FDU) cleaves 1 μg of lambda DNA in 5 minutes 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 PfoI is formulated to digest up to:

  • 1 µg of lambda DNA 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 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 5 5 5 3 65°C, 5 min 16

Methylation Effects

  • Dam: may overlap - cleavage impaired.
  • Dcm: may overlap - blocked.
  • CpG: may overlap - blocked.
  • EcoKI: never overlaps - no effect.
  • EcoBI: never overlaps - no effect.
Methylation type Sequence Cleavage effect
Dam(GATC) 5'...TCCNGGm6A TC...3'
3'...AGGNCC Tm6AG...5'
Impaired
Dcm(CCWGG) 5'...TCm5CA GGA...3'
3'...AG GTm5CCT...5'
Blocked
CpG 5'...TCm5C GGGA...3'
3'...AG Gm5CCCT...5'
Blocked

Number of recognition sites in DNA molecules

Lambda ΦX174 M13mp18/19 pBR322 puc18/19 pUC57
15 0 0 1 1 1
pTZ19R/U pTZ57R pBluescriptIIKS(±) pBluescriptIISK(±) pACYC177 pACYC184
0 0 0 0 1 1

New sites generated by ligation

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
T^CCNGGA TCCNGGCCNGGA
  • [Bme1390I (ScrFI)/FastDigest ScrFI (Bme1390I)]
  • [BssKI]
  • BsuRI (HaeIII)/FastDigest HaeIII (BsuRI)
  • CviJI
T^CCAGGA TCCAGGCCAGGA
  • [Bme1390I (ScrFI)/FastDigest ScrFI (Bme1390I)]
  • [BssKI]
  • BsuRI (HaeIII)/FastDigest HaeIII (BsuRI)
  • CviJI
  • [EcoRII]
  • [MvaI (BstNI)/FastDigest MvaI]
T^CCCGGA TCCCGGCCCGGA
  • [BcnI (NciI)/FastDigest NciI (BcnI)]
  • [Bme1390I (ScrFI)/FastDigest ScrFI (Bme1390I)]
  • BmgT120I
  • [BssKI]
  • BsuRI (HaeIII)/FastDigest HaeIII (BsuRI)
  • Cfr13I (Sau96I)/FastDigest Sau96I (Cfr13I)
  • CviJI
  • [HpaII/FastDigest HpaII]
  • [MspI (HpaII)/FastDigest MspI]
T^CCGGGA TCCGGGCCGGGA
  • [BcnI (NciI)/FastDigest NciI (BcnI)]
  • [Bme1390I (ScrFI)/FastDigest ScrFI (Bme1390I)]
  • BmgT120I
  • [BssKI]
  • BsuRI (HaeIII)/FastDigest HaeIII (BsuRI)
  • Cfr13I (Sau96I)/FastDigest Sau96I (Cfr13I)
  • CviJI
  • [HpaII/FastDigest HpaII]
  • [MspI (HpaII)/FastDigest MspI]
T^CCTGGA TCCTGGCCTGGA
  • [Bme1390I (ScrFI)/FastDigest ScrFI (Bme1390I)]
  • [BssKI]
  • BsuRI (HaeIII)/FastDigest HaeIII (BsuRI)
  • CviJI
  • [EcoRII]
  • [MvaI (BstNI)/FastDigest MvaI]

Obsah balení

20 rxns

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