Team:WashU/Week6

Digest To Check Construct

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.

Redigest and Gel Run

LUCAS!!! BRIAN!!!!!

Kan/C Plates

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.

Check Glycerol Stock

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.

YLC Color Constructs

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