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==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). | |||
==Primer design== | ==Primer design== | ||
We spent the afternoon designing primers. See [[IGEM:Harvard/2006/ | We spent the afternoon designing primers. See [[IGEM:Harvard/2006/Cyanobacteria_Literature#Primer_Design | our primer design section]]. | ||
==More PCC 7942== | ==More PCC 7942== | ||
We asked Alain to order a new stock of PCC 7942 for us, from the [http://www.attc.org ATCC]. | We asked Alain to order a new stock of PCC 7942 for us, from the [http://www.attc.org ATCC]. | ||
Latest revision as of 09:01, 10 August 2006
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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).
Primer design
We spent the afternoon designing primers. See our primer design section.
More PCC 7942
We asked Alain to order a new stock of PCC 7942 for us, from the ATCC.
