From 2012.igem.org
Team:Technion/12_September_2012
Ilya
- Repeated plate reader for tac+RS+mCherry and tac+RBS+mCherry in pCP. Got similar results to previous experiments. Constitutive expression of mCherry, without any effect of theophylline concentrations. Moreover, the tac+RBS+mCherry gave lower readings than tac+RS+mCherry, in contrast to expectations and one previous experiment. After talking to Roee, we decided to summarize the results along with some calculations and assumptions for the failure of the riboswitch. We will still submit this part to the registry, along with all of our results.
- Prepared a 0.4% gel for the gibson attempts on the phage. Unfortunately, we didn't have time to do the experiments today.
- Began setting up the restrictions for characterizing the MCS plasmid. Shahar and Hila will finish the digestions and run the results on a gel.
Inbal
-Colony PCR for the whole part- pTetO+m-cherry+eT7+terminator within pSB1AK3 (let's call it In1, for my convenience)- I picked randomly 5 colonies, The PCR didn't yield any results.
-I PCRed again pTetO+m-cherry (for the T3 cloning), one sample did actually was amplified, but the other- with the same conditions and PCR and equipment, wasn't amplified (the wonder of science!).
- I minipreped In1 from yestreday and the concentration is 202.5 ng/ul.
- I redid a colony PCR for In1 (again picked 5 colonies by random) but didn't get any good results (not the expected band -3600bp).
Asaf
I need to insert the long Riboswitch+polymerase genes to Psb1c3 plasmid.I minipreped the strain that contain the Psb1c3 with the MCS.
I got good DNA concentrations (75.5-120.5 ng/ul).
I cut the Psb1c3 with the MCS with EcoRI and PstI restriction enzymes.
I ran the cut Psb1c3 on gel for extraction. I got the expected 2kb bands.
Shahar cleaned the DNA from the gel I extracted.
Hila
Lior
Noa
Evgeni
Shahar
Rachel
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