Team:Groningen/Labwork/11 September 2013

From 2013.igem.org

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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 <i>amyE<i/> locus.
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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 <i>amyE</i> locus.
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<h2>Sebas</h2>
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<h2>Claudio</h2>
S5 was ligated into pSB1C3-S1-S5 and pSB1C3-S2-S5.
S5 was ligated into pSB1C3-S1-S5 and pSB1C3-S2-S5.
<br>The ligation product was transformed into <i>E. coli</i> DH5&alpha; and plated on LB + Cm.
<br>The ligation product was transformed into <i>E. coli</i> DH5&alpha; and plated on LB + Cm.

Revision as of 10:27, 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%.