MAEIII-RO
Roche
Mae III
from Methanococcus aeolicus PL-15/H
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About This Item
UNSPSC Code:
12352204
biological source
bacterial (Methanococcus aeolicus PL-15/H)
form
solution
packaging
pkg of 250 U (10822248001 [1 - 5 U/μl])
pkg of 50 U (10822230001 [1 - 5 U/μl])
manufacturer/tradename
Roche
parameter
55 °C optimum reaction temp.
storage temp.
−20°C
General description
Mae III is a restriction enzyme which recognizes the sequence ↓GTNAC and generates fragments with 5′-cohesive termini.
Specificity
Recognition sites: GTNAC
GTNAC
Restriction site: ↓GTNAC
↓GTNAC
Heat inactivation: There is no information available whether or not Mae III can be heat inactivated.
GTNAC
Restriction site: ↓GTNAC
↓GTNAC
Heat inactivation: There is no information available whether or not Mae III can be heat inactivated.
Application
Mae III has been used as a restriction enzyme for PCR products.
Quality
Absence of nonspecific endonuclease activities
1μg λDNA is incubated for 16hours in 50μl incubation buffer with an excess of Mae III. The number of enzyme units which do not change the enzyme-specific pattern is stated in the certificate of analysis.
Absence of exonuclease activity
Approximately 5μg [3H] labeled calf thymus DNA are incubated with 3μl Mae III for 4hours at +37°C in a total volume of 100μl 50mM Tris-HCl, 10mM MgCl2, 1mM Dithioerythritol, pH approximately 7.5. Under these conditions, no release of radioactivity is detectable, as stated in the certificate of analysis.
1μg λDNA is incubated for 16hours in 50μl incubation buffer with an excess of Mae III. The number of enzyme units which do not change the enzyme-specific pattern is stated in the certificate of analysis.
Absence of exonuclease activity
Approximately 5μg [3H] labeled calf thymus DNA are incubated with 3μl Mae III for 4hours at +37°C in a total volume of 100μl 50mM Tris-HCl, 10mM MgCl2, 1mM Dithioerythritol, pH approximately 7.5. Under these conditions, no release of radioactivity is detectable, as stated in the certificate of analysis.
DNA Profile
Number of cleavage sites on different DNAs
- λ: 156
- φX174: 17
- Ad2: 118
- M13mp7: 25
- pBR322: 17
- pBR328: 18
- pUC18: 11
- SV40: 14
Unit Definition
One unit is the enzyme activity that completely cleaves 1 μg λDNA in one hour at +55 °C in a total volume of 25 μl (1x) special Mae I incubation buffer.
Analysis Note
Compatible ends
Mae III ends are compatible with ends generated by Bst E II.
Isoschizomers
The enzyme has no known isoschizomers.
Methylation sensitivity
There is no evidence that the enzyme is inhibited by methylation.
SuRE/Cut Buffer System
The buffer in bold is recommended for optimal activity
Incubation temperature
+55°C
Unit definition
One unit is the enzyme activity that completely cleaves 1μg λDNA in 1 hour at +55°C in a total volume of 25μl special incubation buffer.
Heat inactivation
No information available.
Ligation and recutting assay
Mae III fragments obtained by complete digestion of 1μg λDNA are ligated with 1U T4 DNA Ligase in a volume of 10μl by incubation for 16 hours at +4°C in 66mM Tris-HCl, 5mM MgCl2, 5mM Dithiothreitol, 1mM ATP, pH 7.5 (at +8°C) resulting in >95% recovery of 1μg λDNA fragments.
Subsequent re-cutting with Mae III yields >95% of the typical pattern of λDNA × Mae III fragments.
Mae III ends are compatible with ends generated by Bst E II.
Isoschizomers
The enzyme has no known isoschizomers.
Methylation sensitivity
There is no evidence that the enzyme is inhibited by methylation.
SuRE/Cut Buffer System
The buffer in bold is recommended for optimal activity
- A: 0-10%
- B: 10-25%
- H: 10-25%
- L: 0-10%
- M: 0-10%
Incubation temperature
+55°C
Unit definition
One unit is the enzyme activity that completely cleaves 1μg λDNA in 1 hour at +55°C in a total volume of 25μl special incubation buffer.
Heat inactivation
No information available.
Ligation and recutting assay
Mae III fragments obtained by complete digestion of 1μg λDNA are ligated with 1U T4 DNA Ligase in a volume of 10μl by incubation for 16 hours at +4°C in 66mM Tris-HCl, 5mM MgCl2, 5mM Dithiothreitol, 1mM ATP, pH 7.5 (at +8°C) resulting in >95% recovery of 1μg λDNA fragments.
Subsequent re-cutting with Mae III yields >95% of the typical pattern of λDNA × Mae III fragments.
Activity in PCR buffer: 0%
Relative activity in PCR mix (Taq DNA Polymerase buffer) is 0%. The PCR mix contained λDNA, 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.
Relative activity in PCR mix (Taq DNA Polymerase buffer) is 0%. The PCR mix contained λDNA, 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.
Other Notes
For life science research only. Not for use in diagnostic procedures.
Kit Components Only
Product No.
Description
- Enzyme Solution
- Special incubation buffer for Mae III
Storage Class Code
12 - Non Combustible Liquids
WGK
WGK 1
Flash Point(F)
does not flash
Flash Point(C)
does not flash
Regulatory Information
含少量动物源组分生物产品
常规特殊物品
Certificates of Analysis (COA)
Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.
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