Team:Groningen/Notebook/Wetwork 6August2012

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Emeraldo
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Another attempt on inserting the terminator with biobrick suffix and prefix inside pSac-Cm MCS. Terminator+prefix and suffix PCR, cut both original pSac-Cm and PCR product with EcoRI and HindIII, ligation reaction, and then E.coli DH5alpha transformation.
 
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Tom
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PCR alsT (150, 250, 300, 500) + GFP construct with taq-polymerase.
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<body><br><br><br>
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Restriction analysis for alsT (150, 250, 300, 500) + GFP with ecoRI.
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<p class="margin">
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Emeraldo<br>
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Renske
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<br>
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Another attempt on inserting the terminator with biobrick suffix and prefix inside pSac-Cm MCS. Terminator+prefix and suffix PCR, cut both original pSac-Cm and PCR product with EcoRI and HindIII, ligation reaction, and then E.coli DH5alpha transformation.<br>
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Counted TAMC plates + analyzed results:
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<br>
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Tom<br>
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<br>
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PCR alsT (150, 250, 300, 500) + GFP construct with taq-polymerase.<br>
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<html>
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Restriction analysis for alsT (150, 250, 300, 500) + GFP with ecoRI.<br>
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<div style="position:absolute; left: 100px; top: 500px; z-index:-1;">
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<br>
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Renske<br>
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<br>
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Counted TAMC plates + analyzed results:<br>
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<br>
<img src="https://static.igem.org/mediawiki/2012/b/b5/Groningen_RR_20120806_TAMC.png" width="450" height="320">
<img src="https://static.igem.org/mediawiki/2012/b/b5/Groningen_RR_20120806_TAMC.png" width="450" height="320">
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Arjan<br>
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<br>
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GC-MS<br>
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<br>
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Did liquid injection of rotten meat volatiles using methanol as organic solvent on the HP-5 column only. This methode yielded a spectra with multiple peaks and database reference match. <br>
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<br>
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<A HREF="http://www.imik.org/wettelijke_context/Europese_hygienerichtlijn_en_microbiologische_criteria.pdf"><FONT COLOR=#ff6700>reference</FONT></A><br>
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<br>
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Renske<br>
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<br>
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Primer design for positivie feedback loop (with Nisa)<br>
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<br>
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I PlacI<br>
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a) Prefix + PLacI_Forward<br>
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b) homology with pRe_F + PLacI_Rv<br>
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c) homology with pLuxR + PLuxR_Rv <br>
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<br>
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II P_Activator<br>
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a) homology pLac + PRE_F<br>
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b) homology pLac _ pLuxR<br>
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c) homology cII with pRE_R<br>
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d) homology LuxI + pLUxR_R<br>
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<html>
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<br>
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<A HREF="http://www.imik.org/wettelijke_context/Europese_hygienerichtlijn_en_microbiologische_criteria.pdf"><FONT COLOR=#ff6700>reference</FONT></A>
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III Activator<br>
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</html>
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a) homology with pRE + CII_F<br>
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b) homolgy with pLuxI + LuxI_F/ GFP + CII_R/GFP+LuxI_R<br>
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<br>
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Renske
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IV GFP<br>
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a) CII + GFP_F<br>
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Primer design for positivie feedback loop (with Nisa)
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b) LuxI + GFP_R<br>
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c) suffix _ GFP_R<br>
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I PlacI
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<br>
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a) Prefix + PLacI_Forward
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b) homology with pRe_F + PLacI_Rv
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c) homology with pLuxR + PLuxR_Rv  
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II P_Activator
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a) homology pLac + PRE_F
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b) homology pLac _ pLuxR
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c) homology cII with pRE_R
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d) homology LuxI + pLUxR_R
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III Activator
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a) homology with pRE + CII_F
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b) homolgy with pLuxI + LuxI_F/ GFP + CII_R/GFP+LuxI_R
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IV GFP
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a) CII + GFP_F
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b) LuxI + GFP_R
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c) suffix _ GFP_R
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<html>
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<A HREF="https://2012.igem.org/Team:Groningen/Notebook"><FONT COLOR=#ff6700>Back to notebook</FONT></A>
<A HREF="https://2012.igem.org/Team:Groningen/Notebook"><FONT COLOR=#ff6700>Back to notebook</FONT></A>
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<br><br></p></body>
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{{Template:SponsorsGroningen2012}}
{{Template:SponsorsGroningen2012}}

Latest revision as of 13:21, 26 September 2012







Emeraldo

Another attempt on inserting the terminator with biobrick suffix and prefix inside pSac-Cm MCS. Terminator+prefix and suffix PCR, cut both original pSac-Cm and PCR product with EcoRI and HindIII, ligation reaction, and then E.coli DH5alpha transformation.

Tom

PCR alsT (150, 250, 300, 500) + GFP construct with taq-polymerase.
Restriction analysis for alsT (150, 250, 300, 500) + GFP with ecoRI.

Renske

Counted TAMC plates + analyzed results:




Arjan

GC-MS

Did liquid injection of rotten meat volatiles using methanol as organic solvent on the HP-5 column only. This methode yielded a spectra with multiple peaks and database reference match.

reference

Renske

Primer design for positivie feedback loop (with Nisa)

I PlacI
a) Prefix + PLacI_Forward
b) homology with pRe_F + PLacI_Rv
c) homology with pLuxR + PLuxR_Rv

II P_Activator
a) homology pLac + PRE_F
b) homology pLac _ pLuxR
c) homology cII with pRE_R
d) homology LuxI + pLUxR_R

III Activator
a) homology with pRE + CII_F
b) homolgy with pLuxI + LuxI_F/ GFP + CII_R/GFP+LuxI_R

IV GFP
a) CII + GFP_F
b) LuxI + GFP_R
c) suffix _ GFP_R

Back to notebook

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