Team:UNIPV-Pavia/Calendar/July/settimana3

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Revision as of 13:39, 20 July 2010

JULY: WEEK 3

July, 12th

Gel run/cut of phasins amplified via PCR

July, 13th

July, 14th

July, 15th

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)

week 3

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 [[Image:UNIPV10_creening_ligations_I17-I19_I7-8-104C5.jpg|thumb|300px|center|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)