Team:Groningen/Labwork/11 September 2013

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Revision as of 11:52, 11 September 2013

Sebas

Checked sequenced plasmids. BBa_K1085014+GFPdsm and BBa_K1085014+GFP0840 had both the promoter orientated in the right direction.
There were no mutations.

BBa_K1085014+P-RFP, only one primer bound in the wrong direction, implying that the promoter is inserted in the wrong direction. The other primer didn't bound at all.

checked colonies stabbed on LB/starch/cm plates, with lugols solution. No halo's were visible implying that the B. subtilis colonies have an insert in the amyE locus.

Claudio

S5 was ligated into pSB1C3-S1-S5 and pSB1C3-S2-S5.
The ligation product was transformed into E. coli DH5α and plated on LB + Cm.

The plates from yesterday (pSB1C3-S16-S3 and pSB1C3-S16-S9) showed around one hundred colonies.
ColonyPCR was performed on 4 colonies per plate using VF2 and VR primers (annealing temperature 58°C).
The PCR samples were checked on agarose gel 0.8%.

Colony C from pSB1C3-S16-S3 and colonies C and D from pSB1C3-S16-S9 were positive candidates. Indeed the gel showed bands at the expected height (1219 bps).
Colony C from pSB1C3-S16-S3 and colonies C from pSB1C3-S16-S9 were inoculated in LB + Cm (Sander).

@@ Line 40: / Line 40: @@
 <br>The PCR samples were checked on agarose gel 0.8%.
 <br><img src="https://static.igem.org/mediawiki/igem.org/0/0a/ColonyPCR-pSB1C3-SS.png" width="50%" >
-<br>Colony C from pSB1C3-S16-S3 and colonies C and D from pSB1C3-S16-S9 were positive candidates.
+<br>Colony C from pSB1C3-S16-S3 and colonies C and D from pSB1C3-S16-S9 were positive candidates. Indeed the gel showed bands at the expected height (1219 bps).
 <br>Colony C from pSB1C3-S16-S3 and colonies C from pSB1C3-S16-S9 were inoculated in LB + Cm (<b>Sander</b>).