Team:Caltech/Week 6

From 2011.igem.org

(Difference between revisions)
Line 12: Line 12:
Retransform pSB3K5 into Xl-10s<br/>
Retransform pSB3K5 into Xl-10s<br/>
===Results===
===Results===
-
Gels show mostly expected single bands<br/><br/>
+
Gels show mostly expected single bands<br/><br/><br/><br/><br/>
==July 19==
==July 19==
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/>

Revision as of 21:25, 20 July 2011


Caltech iGEM 2011



Home

Project

Data

Parts

Team

Notebook

Biosafety

Human Impact

References

Support

July 18

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
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

Gels show mostly expected single bands




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


Retrieved from "http://2011.igem.org/Team:Caltech/Week_6"