1. Quorum Sensing
Week 5 Plans

2. J-T
OHHL
GFP R I R

3. Quorum or Growth?

4. tetR - Plan I
OHHL R
GFP
tetR
aTc
tetR

5. 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

6. RFP - Plan
RFP
GFP

7. 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

8. 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