. Rachel and Russell ran their RFP/eCFP/AraC isolated plasmids on an agarose gel to see if they were there. The results of the gel seemed unusual.
+
Russell and Rachel ran their isolated plasmids on an agarose gel and found that the bands given were not in-keeping with what they expected. They expected the RFP plasmid to be the largest, eCFP the next largest, and then the two AraC plasmids to be smaller yet but the same size as one another; the results showed RFP and AraC2 to be equal to one another and the largest of the four, while eCFP and AraC1 were shown to be equal to one another and the smallest of the four. It is possible that some plasmids were in their super-coiled states and others were not resulting in the abnormal results and it was concluded that further experiments were needed.
Russell and Rachel ran their isolated plasmids on an agarose gel and found that the bands given were not in-keeping with what they expected. They expected the RFP plasmid to be the largest, eCFP the next largest, and then the two AraC plasmids to be smaller yet but the same size as one another; the results showed RFP and AraC2 to be equal to one another and the largest of the four, while eCFP and AraC1 were shown to be equal to one another and the smallest of the four. It is possible that some plasmids were in their super-coiled states and others were not resulting in the abnormal results and it was concluded that further experiments were needed.
Day 2
. Rachel and Russell calculated the expected length of plasmid and found that the plasmids on the agarose gel were completely different to expected. Decided to carry out a restriction digest of the plasmids to remove the inserts and then run those on a gel to find out if they are as expected. After a nanodrop we found that there was a very small amount of DNA and likely too little to work with; mini-prepped the second samples for each biobrick and nanodropped them.
. Rachel transformed new E. coli with the original biobricks in order to produce a new stock of DNA as we had left gaps between using the bacteria and DNA previously which could account for the very low levels of DNA we had experienced in the nanodrops.
Day 3
. Rachel checked the transformed bacteria and found that some had grown but others hadn't grown quite as much; left for a few more hours to grow before inoculating.
. Russell carried out a restriction digest of each isolated plasmid hopefully containing the RFP, eCFP or AraC biobricks. He did one digest of each plasmid with just EcoR1 to linerarise the plasmid, and one digest of each plasmid with EcoR1 + Pst1 to cut out the insert. These digested plasmids will then be run on an agarose gel in order to validate the DNA that are in the biobricks due to the unusual results previously; if they are fine then the biobricks transformed into E. coli by Rachel will be used to produce new biobricks and carry out quantitative experiments.
"
