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Temperature equilibration took area in much less than five minutes. In two consecutive PCR reactions a I-SceI restriction site and a deletion of 658 bp have been developed. Primers 1001 and 1002 with complementary finishes which includes the recognition site of meganuclease I-SceI, one thousand and 1003 had been utilized. With pCLF3 as template and the talked about primers two fragments correspond