Team:UNIPV-Pavia/Calendar/July/settimana3

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Revision as of 13:25, 26 July 2010

JULY: WEEK 3

July, 12th

Phasins PhaP1 and PhaP2 were sequenced, but none of them has a clear chromatogram, so we decided to sequence the PCR results.

Gel run/cut of phasins amplified via PCR

Phasins were gel run/cut and the samples were prepared to be sequenced.

Inoculum of

<partinfo>BBa_J23101</partinfo>
<partinfo>BBa_J23105</partinfo>
<partinfo>BBa_J23106</partinfo>

for tomorrow ligations.

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/I10-4C5

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

@@ Line 13: / Line 13: @@
 <html><p align="center"><font size="4"><b>JULY: WEEK 3</b></font></p></html><hr><br>
 ==July, 12th==
+Phasins PhaP1 and PhaP2 were sequenced, but none of them has a clear chromatogram, so we decided to sequence the PCR results.
 [[Image:UNIPV10_phasins_PCR.jpg|thumb|200px|center|Gel run/cut of phasins amplified via PCR]]
+Phasins were gel run/cut and the samples were prepared to be sequenced.
+Inoculum of
+*<partinfo>BBa_J23101</partinfo>
+*<partinfo>BBa_J23105</partinfo>
+*<partinfo>BBa_J23106</partinfo>
+for tomorrow ligations.
 ==July, 13th==