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Digestions, ligations, transformations. | Digestions, ligations, transformations. | ||
August 2
Colony PCR to verify ligations. Also some transformations.
August 3
Verifying parts using GFP and making liquid cultures from PCR- and GFP-confirmed colonies.
August 4
We isolated the following assembled parts: F2621-I763020 & R0062-I763020 & F2621-K082014-F2621-I763020 & F2621-I0460-F2621-I763020 & F2621-I0460-K318511 & R0062-I0460-K182102-K082014 & K182102-K082014-R0062-I763020
This day we discovered something horrible... Our sequencing shows that the F2621 part from the registry did not have the R0063 promoter in front of it so at this point we realized we had to redo a lot of our cloning...
August 5
We isolated the following assembled parts: K182102-C0061-B0015 & F2621-I0460
August 6
We isolated the assembled part: K182102-K082014 &
August 8
Digestions, ligations, transformations.
August 10
Digestions, ligations, transformations.
August 11
Yesterday’s gel showed that the digestions were a bit crappy so we redid all of them and then ligated and transformed again. Colony PCR of the transformants from the 10th of August.
August 12
Miniprepped the following parts: R0063-F2621-I0460 & R0063-F2621-I763020 & F2621-E0422-F2621-K082014 & R0063-F2621-K082014 & K176028-I0460 Digestions were started to then be ligated and used to transform E. coli.
August 13
PCR screen of the ligations from August 12th. Also minipreps for sequencing of K182102-K082014-I763020 & R0063-F2621-I763020 & R0063-F2621-I0460 & R0063-F2621-K082014 & I0460-K176028 We also made liquid cultures from the colonies that got PCR-confirmed.
August 14
Miniprepped the following parts: R0063-F2621-K082014-R0063-F2621-I763020 & R0063-F2621-I763020-R0063-F2621-K082014 More digestions and ligations and transformations.
August 15
Colony pcr from yesterdays’ ligations.
August 16
Miniprepped the following parts: J23105-P0312 & K176028-I0460-K182102-K082014-R0062-I763020
August 17
Digestions, ligations, transformations.
August 18
Colony PCR
August 23
Performing a double ligation using R0040, J24675 & a linearized backbone.
August 25
Miniprepped R0040-J24675
August 30
Digestions, ligations, transformations.
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