JULY: WEEK 3
July, 12th
Gel run/cut of phasins amplified via PCR
July, 13th
July, 14th
Quality control for HELPER and CRIM plasmids
July, 15th
Quality control for pBHR68 and CRIM plasmids
July, 16th
Screening ligations I11-I16
Screening ligations I17-I19, I7/I8/I104C5
Since it's impossible to isolate the CRIM plasmid <partinfo>BBa_J72007</partinfo> we created a pseudo CRIM plasmid to check that not pir strains aren't able to propagate it.
We called it "THE RING" (RING) and we built it from our I5 (Cm resistance - <partinfo>BBa_P1004</partinfo> - + TT - <partinfo>BBa_B0015</partinfo> - + R6K ori - <partinfo>BBa_J61001</partinfo>) in <partinfo>pSB1A2</partinfo>:
- Digestion of I5 XbaI-PstI
- Gel extraction
- Quantification with Nanodrop (15ng/ul)
- Ligation of X and P sites each other (used 2ul of DNA)
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