Quorum Sensing Week 5 Plans

J-T OHHL GFP R I R

Quorum or Growth?

tetR - Plan I OHHL R GFP tetR aTc tetR

tetR - Schedule Date tetR Monday Tuesday Wednesday Thursday Friday Colony PCR transformed [R00XX][P0X40] Send promoters for sequencing Grow liquid cultures Tuesday Miniprep, digest, dephosphorylate, clonewell, and ligate [R00XX][P0X40] with [J23039][T9002] constructs Transform final constructs into BL21 Wednesday Colony PCR [R00XX][P0X40][J23039][T9002] Send for sequencing Grow liquid cultures of [R00XX][P0X40][J23039][T9002] Thursday Set up fluorescence plate reader of final constructs Friday Observe results

RFP - Plan RFP GFP

RFP – Schedule Date RFP tetR Monday Tuesday Wednesday Thursday Friday Miniprep J04450 Digest + dephosphorylate Ligate Transform into BL21 cells Colony PCR transformed [R00XX][P0X40] Send promoters for sequencing Grow liquid cultures Tuesday Colony PCR Sequence Miniprep, digest, dephosphorylate, clonewell, and ligate Transform final constructs into BL21 Wednesday Set up fluorescence plate reader of J50-J39-T02 Send for sequencing Thursday Observe results Set up fluorescence plate reader of final constructs Friday

OD vs. Fluorescence Date OD RFP tetR Monday Tuesday Wednesday Thursday Set up overnight plate reader for OD Miniprep J04450 Digest + dephosphorylate Ligate Transform into BL21 cells Colony PCR Send promoters for sequencing Grow liquid cultures Tuesday Set up overnight plate reader for fluorescence Sequence Miniprep, digest, dephosphorylate, clonewell, and ligate Transform Wednesday Set up fluorescence plate reader of J50-J39-T02 Send for sequencing Thursday Observe results Set up fluorescence plate reader of final constructs Friday