Sequence of PCRing
We've decided to lift the whole KaiABC segment out of cyanobacteria first, then break it into pieces later (KaiA operon, KaiA coding region, KaiBC operon, KaiBC coding region), since it's easier to work with shorter lengths of DNA.
There are 3 BioBrick restriction sites in KaiABC which we will need to modify with site-directed mutagenesis before inserting the sequence into a plasmid. This step will come immediately after or concurrent with the first PCR (check on this? -Jeff).
We spent the afternoon designing primers. See our primer design section.