Erase-a-Base™ System

The Erase-a-Base™ System is designed for the rapid construction of plasmid or M13 subclones containing progressive unidirectional deletions of any inserted DNA. The system is based on the procedure developed by Henikoff, in which exonuclease III (Exo III) is used to specifically digest insert DNA from a 5´ protruding or blunt end restriction site. The adjacent sequencing primer binding site is protected from digestion by a 4-base 3´ overhang restriction site or by an α-phosphorothioate-filled end.

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The Erase-a-Base™ System is designed for the rapid construction of plasmid or M13 subclones containing progressive unidirectional deletions of any inserted DNA. The system is based on the procedure developed by Henikoff, in which exonuclease III (Exo III) is used to specifically digest insert DNA from a 5´ protruding or blunt end restriction site. The adjacent sequencing primer binding site is protected from digestion by a 4-base 3´ overhang restriction site or by an α-phosphorothioate-filled end.

Features - Benefits

• Rapid: Construction of nested deletions from plasmid or M13 clones are rapid. Construction is complete in a few hours.
• Efficient: Produce deletion sets spanning several kilobases.

References

1. Henikoff, S. (1984) Gene 28, 351–9.
2. Putney, S.D. et al. (1981) Proc. Natl. Acad. Sci. USA 78, 7350–4.

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