Analysis Note

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.

Compatible endsMae III ends are compatible with ends generated by Bst E II.IsoschizomersThe enzyme has no known isoschizomers.Methylation sensitivityThere is no evidence that the enzyme is inhibited by methylation.SuRE/Cut Buffer SystemThe 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°CUnit definitionOne 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 inactivationNo information available.Ligation and recutting assayMae 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.

Application

Mae III has been used as a restriction enzyme for PCR products.

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

General description

Mae III is a restriction enzyme which recognizes the sequence ↓GTNAC and generates fragments with 5′-cohesive termini.

Other Notes

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

Quality

Absence of nonspecific endonuclease activities1µg 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 activityApproximately 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.

Specificity

Recognition sites: GTNACGTNACRestriction site: ↓GTNAC↓GTNACHeat inactivation: There is no information available whether or not Mae III can be heat inactivated.

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.