| Identification | Back Directory | [Name]
SPH I | [CAS]
85270-15-1 | [Synonyms]
APA I
SPH I
Einecs 286-569-8
SPH I RESTRICTION ENZYME
APA I RESTRICTION ENZYME
RESTRICTION ENDONUCLEASE APA I
restriction endonuclease Sph I
RESTRICTION ENDONUCLEASES APA I
RESTRICTION ENDONUCLEASES SPH I
apa i from acetobacter pasteurianus
sph i from streptomyces phaeochromogenes
Nuclease, restriction endodeoxyribo-, SphI
restriction endonuclease apa I from*acetobacter P
RESTRICTION ENDONUCLEASE SPH I FROM*STRE PTOMYCES
RESTRICTION ENDONUCLEASE APA I FROM*ACET OBACTER PAS | [EINECS(EC#)]
286-569-8 | [MDL Number]
MFCD00166621 |
| Hazard Information | Back Directory | [General Description]
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.
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| Company Name: |
Sigma-Aldrich
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| Tel: |
021-61415566 800-8193336 |
| Website: |
https://www.sigmaaldrich.cn |
| Company Name: |
Creative Enzymes
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| Tel: |
1-516-855-7709 |
| Website: |
www.creative-enzymes.com |
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