THERMOPHILIC STRAND DISPLACEMENT AMPLIFICATIONS USING AVAI

摘要

The mesophilic restriction endonuclease AvaI and its isoschizomers derived from Anabaena and Anabaenopsis have been found to support highly efficient amplificat ion of target nucleic acid sequences in thermophilic Strand Displacement Amplification (tSDA). The maximum amplification factors observed occurred at temperatures which were previ ously reported to result in reduced enzyme activity. As AvaI is an isoschizomer of Bso BI (a thermophilic restriction endonuclease commonly used in tSDA), the invention prov ides an alternative enzyme which is more readily available than BsoBI and does not requi re the redesign of SDA amplification primers previously constructed for use in the BsoB I system.