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Team:Caltech/Notebook/Coliroid
From 2012.igem.org
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Coliroid NotebookWe grew colonies of R0082 and E1010 and Mg1655 strains overnight. Back to Coliroid Notebook Calendar 7/3/12 We made 3 plates containing Mg1655, putting 1 microliter of Mg1655 on 1 plate, 10 microliters on the second plate, and 100 microliters on the third plate, in order to figure out which one would make the best looking plate of red E. coli. We also miniprepped samples of R0028 and E1010. Back to Coliroid Notebook Calendar 7/3/12 We double digested R0082, lacZ, and E1010 (RFP). We ran the digested samples on a gel along with undigested samples of each to make sure the digests worked. The lacZ and the E1010 digests were successful, but the gel results on the R0082 were inconclusive. We also transformed pSBC13 into Mg1655, but the LB was contaminated so we did not plate the culture. Back to Coliroid Notebook Calendar 7/3/12
We started another culture for M1655 since our previous culture was made with contaminated LB. Then we re-digested the R0082 and ran a gel comparing the digested R0082 with the undigested. The digest was successful, so tested the concentrations of DNA in the R0082, the E1010, and the lacZ samples with the Nanodrop. There was not a high enough concentration of the E1010 DNA, so we could not use it in ligation. We ligated the R0082 with the E1010 and plated the plasmid. Also, we ordered primers for mCherry-AAV, mCherry-LVA, and mCherry.
We found that there was growth on the ligation plate and no growth on the negative control plate, which was good. We checked to make sure that the ligation of R0082 and E1010 worked by running a PCR of 6 different colonies on the ligation plate as well as a sample of the undigested R0082 and then running a gel of these things. Back to Coliroid Notebook Calendar |
