Mae III

Кат. номер

10822248001

10822230001

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.
Mae III has been used as a restriction enzyme for PCR products.
Quality
Absence of nonspecific endonuclease activities
1µg DNA 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

• : 156
• X174: 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 DNA 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 DNA 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 DNA 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 DNA fragments.
Subsequent re-cutting with Mae III yields >95% of the typical pattern of DNA x Mae III fragments.
Activity in PCR buffer: 0%
Relative activity in PCR mix (Taq DNA Polymerase buffer) is 0%. The PCR mix contained 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.
For life science research only. Not for use in diagnostic procedures.

Quality Level	100,
form	solution
packaging	pkg of 250 U (10822248001 [1 - 5 U/µl]), pkg of 50 U (10822230001 [1 - 5 U/µl])
Торговая марка	Roche
parameter	55 °C optimum reaction temp.
shipped in	dry ice
storage temp.	20°C

RIDADR

NONH for all modes of transport