FastDigest® Acc65I

Kód produktu: FD0904 Kód výrobce: FD0904 Kód dodavatele: {64C74208-B479-4739-890C-78EFF0C81BE7} Výrobce: UAB Fermentas
1 219,68 Kč
1 008,00 Kč bez DPH
do týdne
100 react

5'...GG T A C C...3'
3'...C C A T GG...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 FastDigest BanI (BshNI), FastDigest BsiWI (Pfl23II), FastDigest Bsp1407I, FastDigest TatI, BanI, BshNI, Bsp1407I, Pfl23II, TatI.
Formulation 1 μL of enzyme (1 FDU) cleaves 1 μg of lambda DNA/BamHI 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 Acc65I is formulated to digest up to:

  • 1 µg of lambda DNA/BamHI 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 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, minutes 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 minutes 16

Methylation Effects

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


Dcm (CCWGG)

5'...GGTACm5CW GG...3'
3'...CCATG GWm5CC...5'
Impaired


Dcm (CCWGG)

5'...Cm5CW GGTACm5CW GG...3'
3'...G GWm5CCATG GWm5CC...5'
Blocked
CpG 5'...GGTACm5C G...3'
3'...CCATG Gm5C...5'
Impaired
CpG 5'...m5C GGTACm5C G...3'
3'... Gm5CCATG Gm5C...5'
Blocked

Number of recognition sites in DNA molecules

Lambda ΦX174 M13mp18/19 pBR322 puc18/19 pUC57
2 0 1 0 1 1
pTZ19R/U pTZ57R pBluescriptIIKS(±) pBluescriptIISK(±) pACYC177 pACYC184
1 1 1 1 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
G^GTACC
  • BshNI (BanI)/FastDigest BanI (BshNI)* (G^GTACC)
  • Acc65I (Asp718I)/FastDigest Acc65I
  • BshNI (BanI)/FastDigest BanI (BshNI)
  • BspLI (NlaIV)/FastDigest NlaIV (BspLI)
  • Csp6I (CviQI)/FastDigest Csp6I
  • KpnI/FastDigest KpnI
  • RsaI/FastDigest RsaI
  • Bsp1407I (BsrGI)/FastDigest Bsp1407I (T^GTACA)
  • Pfl23II (BsiWI)/FastDigest BsiWI (Pfl23II) (C^GTACG)
  • TatI/FastDigest TatI* (W^GTACA)
  • TatI/FastDigest TatI* (W^GTACT)
  • Csp6I (CviQI)/FastDigest Csp6I
  • RsaI/FastDigest RsaI

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^GTACC GGTACGTACC
  • [Csp6I (CviQI)/FastDigest Csp6I]
  • Eco105I (SnaBI)/FastDigest SnaBI (Eco105I)
  • MaeII
  • Ppu21I (BsaAI)/FastDigest BsaAI (Ppu21I)
  • [RsaI/FastDigest RsaI]
  • SetI
  • TaiI (MaeII)/FastDigest TaiI

Obsah balení

100 rxns

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