MAEIII-RO

Mae III

from Methanococcus aeolicus PL-15/H

• UNSPSC Code: 12352204

Properties

• 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

Description

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.

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.

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

• A: 0-10%
• B: 10-25%
• H: 10-25%
• L: 0-10%
• M: 0-10%

Note: Instead of the SuRE/Cut buffer system, please use the special 2x incubation buffer supplied with the enzyme.

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.

Other Notes

For life science research only. Not for use in diagnostic procedures.

Kit Components Only

• Enzyme Solution
• Special incubation buffer for Mae III

Safety Information

• Storage Class Code: 12 - Non Combustible Liquids
• WGK: WGK 1
• Flash Point(F): does not flash
• Flash Point(C): does not flash