Team:Caltech/Week 6

From 2011.igem.org

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Retransform pSB3K5 into Xl-10s<br/>
Retransform pSB3K5 into Xl-10s<br/>
===Results===
===Results===
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[[File:Gel1_7_9_Caltech.jpg|thumb|left|Colonies 1-6 from Gibson plates; 7 and 8 are reswabs of two colonies. From left to right: DNA ladder,(PNT001; colony 1; vector), P2v, P3v, P4v, P5v, P6v, P7v, P8v, (PNT001; colony 1; insert), P3i, P4i, P5i, P6i]]
+
[[File:Caltech_7_19_Gel_1.jpg|thumb|left|Colonies 1-6 from Gibson plates; 7 and 8 are reswabs of two colonies. From left to right: DNA ladder,(PNT001; colony 1; vector), P2v, P3v, P4v, P5v, P6v, P7v, P8v, (PNT001; colony 1; insert), P3i, P4i, P5i, P6i]]
[[File:Gel2_7_19_Caltech.jpg.jpg|thumb|left|from left to right: DNA ladder, (PNT001; reswab 7; insert), P8i, (16s; colony A; vector), 16Bv, 16Cv, (16s; A; insert), 16Bi, 16Ci]]<br/><br/><br/><br/><br/><br/><br/><br/><br/><br/><br/><br/><br/><br/><br/><br/>
[[File:Gel2_7_19_Caltech.jpg.jpg|thumb|left|from left to right: DNA ladder, (PNT001; reswab 7; insert), P8i, (16s; colony A; vector), 16Bv, 16Cv, (16s; A; insert), 16Bi, 16Ci]]<br/><br/><br/><br/><br/><br/><br/><br/><br/><br/><br/><br/><br/><br/><br/><br/>
==July 19==
==July 19==
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PCR parts for Gibson assembly of pNT001, pNT003 and Joe's positive GFP control<br>
+
PCR parts for Gibson assembly of pNT001, pNT003 and Joe's positive GFP control<br/>
===Results===
===Results===
pSB3K5 plate had 0 colonies.<br/>
pSB3K5 plate had 0 colonies.<br/>

Revision as of 21:16, 20 July 2011


Caltech iGEM 2011



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July 18

Colony PCR of colonies from positive Gibson plates from Friday, for vector and insert
Colony PCR of 16s plasmids, for vector and insert
Gel of short PCR parts to confirm they are being amplified
Pulsed field gel with a larger amount of soil sample DNA
Retransform pSB3K5 into Xl-10s

Results

Colonies 1-6 from Gibson plates; 7 and 8 are reswabs of two colonies. From left to right: DNA ladder,(PNT001; colony 1; vector), P2v, P3v, P4v, P5v, P6v, P7v, P8v, (PNT001; colony 1; insert), P3i, P4i, P5i, P6i
from left to right: DNA ladder, (PNT001; reswab 7; insert), P8i, (16s; colony A; vector), 16Bv, 16Cv, (16s; A; insert), 16Bi, 16Ci
















July 19

PCR parts for Gibson assembly of pNT001, pNT003 and Joe's positive GFP control

Results

pSB3K5 plate had 0 colonies.
PFGE of soil-extracted DNA shows a smear; will do gel extraction of ~40kbp sequences for fosmid kit.

Gibson PCR concentrations

Part Concentration (ng/ul)
R0010 for pNT001 86.4
B0014 for pNT001 128.3
B0014 for pNT003 89.1
promoter for GFP Gibson positive control 51.9
GFP for positive control 130.4
terminator for positive control 82.5


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