The minipreped constructs from Saturday were digested with XbaI and PstI to check if they contained the desired plasmid and insert. Here are the nanodrop results of the potential ligations:
(insert table)
A 1% agarose gel was made to check the digests of the ligation. Here is an image of the gel:
(gel picture)
The ligations were deemed unsuccessful, and most were likely the plasmid backbone with the small self-ligated addition filling in the cut. We will re-digest the insert and plasmid and gel purify a large quantity of DNA to reduce the risk of self-ligation.
LUCAS!!! BRIAN!!!!!
Kanamycin-chloramphenicol plates were made to select for E. coli that take up our construct to produce Saffron from Zeaxanthin and Part:BBa_K395704 which will bring E. coli to Zeaxanthin. Our construct will be Kan-resistant while Part:BBa_K395704 will be C-resistant. The plate will select for the double transformations. The plates were made using the LB-plate protocol on the protocol page.
One of the glycerol stocks was checked by thawing a tube from Saturday on ice and adding .4 mL of it to 3 mL of LB+Amp liquid. The liquid will shake over night at 37°C. The rest of the glycerol stock was refrozen at -80°C.
Since the color constructs are proving difficult, and since we will not have the primers until Friday, we are rehydrating and transforming 4 others constructs to have the colors ready for the YLC project demonstration in one week. The following 4 parts were used to transform cells that will incubate at 37°C overnight: RFP: I13521 GFP: I13522 YFP: 13604 CFP: S03475 75 mL of the LB solution that shook for an hour during transformation was used to plate the cells.
Tuesday, July 3
Thursday, July 5
Friday, July 6
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