SPH I

SPH I Chemische Eigenschaften,Einsatz,Produktion Methoden

• Allgemeine Beschreibung

  Compatible ends
  Apa I ends are not compatible with those generated by any other known restriction enzymes.
  
  Isoschizomers
  Apa I is an isoschizomer to Bsp 120 I and Psp OMI.
  
  Methylation sensitivity
  Apa I is inhibited by 5′-methylcytosine at the sites indicated (*) in the recognition sequence.
  
  Activity in SuRE/Cut Buffer System
  Buffer printed in bold face type is the buffer recommended for optimal activity:
  ABLMH
  100%10-25%50-75%50-75%0-10%
  
  Relative activity in complete PCR mix
  Relative activity in PCR mix (Taq DNA Polymerase buffer) is 100%. The PCR mix contained target DNA, primers, 10 mM Tris-HCl (pH 8.3, +20°C), 50 mM KCl, 1.5 mM MgCl2, 200 μM dNTPs, 2.5 U Taq DNA polymerase. The mix was subjected to 25 amplification cycles. Activity in reaction buffer of Pwo SuperYield DNA Polymerase PCR Mix is 10%. When supplemented with GC-RICH Solution activity is increased to >100%.
  
  Incubation temperature
  +30°C
  
  Unit definition
  One unit is the enzyme activity that completely cleaves 1 μg ? × Hind III fragments in 1 hour at +30°C in a total volume of 25 μl SuRE/Cut Buffer A.
  
  Heat inactivation
  The enzyme can be heat inactivated by incubating it for 15 minutes at +65°C.
  
  Number of cleavage sites on different DNAs
  λAd2SV40φ X174M13mp7M13mp18pBR322pBR328pUC18
  112 1000000
  
  Ligation and recutting assay
  Apa I fragments obtained by complete digestion of 1 μg λ × Hind III fragments are ligated with 1 U T4 DNA Ligase in a volume of 10 μl by incubation for 16 hours at +4°C in 66 mM Tris-HCl, 5 mM MgCl2, 5 mM Dithiothreitol, 1 mM ATP, pH 7.5 (at +20°C) resulting in >95% recovery of 1 μg λ × Hind III fragments.
  Subsequent re-cutting with Apa I yields >95% of the typical pattern of λ × Hind III× Apa I fragments.
  

SPH I Anbieter Lieferant Produzent Hersteller Vertrieb Händler.

Global(63)Suppliers

85270-15-1, SPH I Verwandte Suche: