Team:Northwestern/CFPS
From 2014.igem.org
Cell-Free Protein Synthesis
20140723 Testing our lysates
- data from BL21 (E.Coli) lysates
- out of K12, DH5a, and BL21, BL21 expressed the most fluorescence
- the plasmid used was a construct of superfolder GFP from the Jewett Lab
- conclusion:our lysates are functional
![red: control, blue: BL21 lysate](https://static.igem.org/mediawiki/2014/0/05/072314CFPS.png)
![](https://static.igem.org/mediawiki/2014/6/65/E.coli_lysates.jpg)
![](https://static.igem.org/mediawiki/2014/3/32/E.coli_lysates_2.jpg)
20140801 Making our own mastermix (iMM)!
- RM: Rey's mastermix
- iMM: we made our own according to Rey's recipe
- looks like our mastermix works fine
- conclusion: we will use our own mastermix from now on
- the second part: we made s. rimosus lysates
- we used a sfgfp construct from the Jewett lab
- the lysates seem to express some gfp!
![](https://static.igem.org/mediawiki/2014/5/52/IMMRMBL21.jpg)
![](https://static.igem.org/mediawiki/2014/1/1b/Srsfgfp.jpg)
20140915 All of our lysates
- RBS B0034
- Tried all of our lysates
- green fluorescence indicates mRNA levels of transcription
- red fluorescence indicates protein levels of translation
- conclusion: the data is most likely statistically insignificant
![](https://static.igem.org/mediawiki/2014/9/94/RFP091514.png)
![](https://static.igem.org/mediawiki/2014/c/cf/GFP_091514.png)
20141014 Cell-Free Protein Synthesis Assay with All of our Lysates
- We used RBS No. 5 (B0034) with RFP-Spinach Aptamer
- Displayed here is the first read
- The kinetic portion of the plate reader did not record data in all of our wells
- We will continue doing CFPS experiments
- conclusion: we need more time! Looks like fluorophores were incorrectly added
![](https://static.igem.org/mediawiki/2014/f/f2/Otherlysatesoctober.jpg)
![](https://static.igem.org/mediawiki/2014/4/4a/E.colioctober.jpg)
![](https://static.igem.org/mediawiki/2014/0/00/Spinapt101514.png)
![](https://static.igem.org/mediawiki/2014/c/c0/Jewettlabsfgfp.png)
![](https://static.igem.org/mediawiki/2014/c/cf/RFPE.Coli101514.png)