Cut'N'Survive System Functional Principle Sequences Schedule Jump-or-Die System Functional Principle Sequences Schedule ProSearch System Schedule Functional Principle Sequences Schedule picture gallery 1TreF 56°C: *Number: consecutive numbering on PCR plan *Tre: Abbreviation of gene of interest *F: Forward Primer *56°C: Annealing temperature (4+2 rule) 4TevMutEPR : *Mut: Mutagenesis Primer *E: EcoRI digestion site *P: PstI digestion site *R: Reverse Primer Also: *S: SpeI digestion site Primer Primer overhang with Biobrick digestion sites Digestion site which is to be mutated Template