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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® NdeI

FastDigest® NdeI

Kód produktu: FD0585 Kód výrobce: FD0585 Kód dodavatele: {91B5B0D2-884F-4860-AEB1-2153D6795A48} Výrobce: Life Technologies Czech Republic s.r.o.
FastDigest® NdeI
38 066,60 Kč
31 460,00 Kč bez DPH
do týdne
1000 rxns

5'...C A↓T A T G...3'
3'...G T A T↑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.

Ends FastDigest AseI (VspI), FastDigest BfaI (FspBI), FastDigest Csp6I, FastDigest MseI (SaqAI), FastDigest Tru1I, AseI, BfaI, Csp6I, FspBI, Tru1I, VspI.Isoschizomers Search for commercial isoschizomers using REsearch.
Compatible
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
Recommended Reaction Conditions

1X FastDigest buffer or 1X FastDigest Green buffer at 37°C.
1 µl of FastDigest NdeI 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 > 60 minutes
  • 1 µg of genomic DNA in 30 minutes or 5 µg of genomic DNA in 3 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, 1µg/20µl PCR product, ~0.2 µg/30 µl Genomic DNA, 1 µg/10 µl
37°C 5 5 60 30 3 65°C, 5 min 6

Methylation Effects

  • Dam: never overlaps - no effect.
  • Dcm: never overlaps - no effect.
  • CpG: never overlaps - no effect.
  • EcoKI: never overlaps - no effect.
  • EcoBI: may overlap - no effect.
Methylation type Sequence Cleavage effect
EcoBI(TGA(N)8TGCT)

5'...TGm6A(N)4CATA TGCT...3'
3'...AC T(N)4GTATm6ACGA...5'

 No effect

Number of recognition sites in DNA molecules

Lambda ΦX174 M13mp18/19 pBR322 puc18/19 pUC57
7 0 3 1 1 1
pTZ19R/U pTZ57R pBluescriptIIKS(±) pBluescriptIISK(±) pACYC177 pACYC184
0 0 0 0 0 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
CA^TATG
  • Csp6I (CviQI)/FastDigest Csp6I (G^TAC)
  • FspBI (BfaI)/FastDigest BfaI (FspBI) (C^TAG)
  • FastDigest MseI (SaqAI) (T^TAA)
  • Tru1I (MseI)/FastDigest Tru1I (T^TAA)
  • VspI (AseI)/FastDigest AseI (VspI) (AT^TAAT)
  • FaiI

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
CA^TATG CATATATG
  • [FaiI]

Obsah balení

1000 rxns

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