χρόνος πέρασμα May 10th 2011
- Today we have isolated again the plasmid pBBRMCS5. We have also isolated the plasmid pSB4C5, from which we will amplify a specific region that contains the gene that codifies for the Red Flourescent Protein (RFP) and the respective restriction enzyme sites that makes a Biobrick as such. This region of the plasmid pSB4C5 is the one we wish to introduce to the other plasmids (pBBRMCS5 and pRK404)in order to leave them standarized.
- We have followed the protocol provided by Miguel Ángel Ramírez with some modifications.
-We have skipped the steps 11, 13-20.
-The centrifugations were performed under room temperature instead of 4° Celcius.
-The step 4 was followed by TE 10:1 wash.
-The final step (24) does not say the amount of TE-RNAse to be applied to the extracted plasmid.
-Paz told me to use 20 microliter and let it rest at 37° for one hour prior to run the plasmids in an electrophoresis gel.
-Uriel told us to use 60 microliters (we'll do it next time).
- We ran the electrophoresis gel with the isolated plasmids. The gel contained 15 wells, numbers 1 and 15 had the DNA ladder, numbers 2-6 had the plasmid pBBRMCS5, number 7 contained the control (just the dye), numbers 8-13 had the plasmid pSB4C5, number 14 was empty.
- It should be emphasized that we couldn't see the Matrix movies because we were working all day, we should be rewarded with many iGEMpoints :)