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Team:Northwestern/CFPS
From 2014.igem.org
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| + | <h5>20140801 Making our own mastermix (iMM)!</h5> | ||
| + | <ul> | ||
| + | <li>RM: Rey's mastermix</li> | ||
| + | <li>iMM: we made our own according to Rey's recipe</li> | ||
| + | <li>looks like our mastermix works fine</li> | ||
| + | <li><b>conclusion</b>: we will use our own mastermix from now on</li> | ||
| + | <li>the second part: we made s. rimosus lysates</li> | ||
| + | <li>we used a sfgfp construct from the Jewett lab</li> | ||
| + | <li>the lysates seem to express some gfp!</li> | ||
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Latest revision as of 23:19, 17 October 2014
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
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!
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
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
