Team:KULeuven/18 August 2008

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=== Wet Lab ===
=== Wet Lab ===
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[[image:gel-18-8a.jpg|right|thumb|340px|'''Gel 1''': PCR product of K145151 and K145013]]
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[[image:gel-18-8b.jpg|right|thumb|340px|'''Gel 2''': Digest of B0015 and F1610 with ''Xba'' I]]
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We did some PCRs with ''pfx'' polymerase to prepare the following (new) BioBricks: [http://partsregistry.org/Part:BBa_K145014 K145014] (T7 polymerase with UmuD), [http://partsregistry.org/Part:BBa_K145013 K145013] (antisense LuxI) and [http://partsregistry.org/Part:BBa_K145151 K145151] (ccdB). The templates and primers used were, respectively, PCR of step 1 with 1077/1079, BBa_F1610 with 1100/1101, BBa_P1010 with 1127/1128. Only the PCR of ccdB succeeded. (see '''Gel 1''').
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We made a few digests: [http://partsregistry.org/Part:BBa_F1610 F1610] and [http://partsregistry.org/Part:BBa_B0015 B0015] were cut with ''Eco''RI and ''Xba''I (see '''Gel 2'''), and [http://partsregistry.org/Part:BBa_J23100 J23100]+[http://partsregistry.org/Part:BBa_B0032 B0032] and [http://partsregistry.org/Part:BBa_R0062 R0062]+[http://partsregistry.org/Part:BBa_B0032 B0032] were cut with ''Eco''RI and ''Spe''I.
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We electroporated the following ligated parts: [http://partsregistry.org/Part:BBa_C0040 C0040]+[http://partsregistry.org/Part:BBa_B0015 B0015] , [http://partsregistry.org/Part:BBa_K145015 K145015]+[http://partsregistry.org/Part:BBa_B0015 B0015], [http://partsregistry.org/Part:BBa_C0012 C0012]+[http://partsregistry.org/Part:BBa_B0015 B0015] and [http://partsregistry.org/Part:BBa_C0060 C0060]+[http://partsregistry.org/Part:BBa_B0015 B0015].
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We continued ligating: [http://partsregistry.org/Part:BBa_J23116 J23116]+[http://partsregistry.org/Part:BBa_B0032 B0032] , [http://partsregistry.org/Part:BBa_K145001 K145001]+[http://partsregistry.org/Part:pSB1A2 pSB1A2], [http://partsregistry.org/Part:BBa_R0040 R0040]+[http://partsregistry.org/Part:BBa_B0032 B0032], [http://partsregistry.org/Part:BBa_R0040 R0040]+[http://partsregistry.org/Part:BBa_J23022 J23022], [http://partsregistry.org/Part:BBa_R0084 R0084]+[http://partsregistry.org/Part:BBa_B0032 B0032], [http://partsregistry.org/Part:BBa_K145001 K145001]+[http://partsregistry.org/Part:BBa_B0015 B0015] and ([http://partsregistry.org/Part:BBa_R0084 R0084]+[http://partsregistry.org/Part:BBa_J23022 J23022])+([http://partsregistry.org/Part:BBa_J23109 J23109]+[http://partsregistry.org/Part:BBa_J23032 J23032]). This time, B0032 was first dephosphorylated.
=== Dry Lab ===
=== Dry Lab ===

Latest revision as of 16:27, 11 October 2008

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Contents

Lab Work

Wet Lab

Gel 1: PCR product of K145151 and K145013
Gel 2: Digest of B0015 and F1610 with Xba I

We did some PCRs with pfx polymerase to prepare the following (new) BioBricks: [http://partsregistry.org/Part:BBa_K145014 K145014] (T7 polymerase with UmuD), [http://partsregistry.org/Part:BBa_K145013 K145013] (antisense LuxI) and [http://partsregistry.org/Part:BBa_K145151 K145151] (ccdB). The templates and primers used were, respectively, PCR of step 1 with 1077/1079, BBa_F1610 with 1100/1101, BBa_P1010 with 1127/1128. Only the PCR of ccdB succeeded. (see Gel 1).

We made a few digests: [http://partsregistry.org/Part:BBa_F1610 F1610] and [http://partsregistry.org/Part:BBa_B0015 B0015] were cut with EcoRI and XbaI (see Gel 2), and [http://partsregistry.org/Part:BBa_J23100 J23100]+[http://partsregistry.org/Part:BBa_B0032 B0032] and [http://partsregistry.org/Part:BBa_R0062 R0062]+[http://partsregistry.org/Part:BBa_B0032 B0032] were cut with EcoRI and SpeI.

We electroporated the following ligated parts: [http://partsregistry.org/Part:BBa_C0040 C0040]+[http://partsregistry.org/Part:BBa_B0015 B0015] , [http://partsregistry.org/Part:BBa_K145015 K145015]+[http://partsregistry.org/Part:BBa_B0015 B0015], [http://partsregistry.org/Part:BBa_C0012 C0012]+[http://partsregistry.org/Part:BBa_B0015 B0015] and [http://partsregistry.org/Part:BBa_C0060 C0060]+[http://partsregistry.org/Part:BBa_B0015 B0015].

We continued ligating: [http://partsregistry.org/Part:BBa_J23116 J23116]+[http://partsregistry.org/Part:BBa_B0032 B0032] , [http://partsregistry.org/Part:BBa_K145001 K145001]+[http://partsregistry.org/Part:pSB1A2 pSB1A2], [http://partsregistry.org/Part:BBa_R0040 R0040]+[http://partsregistry.org/Part:BBa_B0032 B0032], [http://partsregistry.org/Part:BBa_R0040 R0040]+[http://partsregistry.org/Part:BBa_J23022 J23022], [http://partsregistry.org/Part:BBa_R0084 R0084]+[http://partsregistry.org/Part:BBa_B0032 B0032], [http://partsregistry.org/Part:BBa_K145001 K145001]+[http://partsregistry.org/Part:BBa_B0015 B0015] and ([http://partsregistry.org/Part:BBa_R0084 R0084]+[http://partsregistry.org/Part:BBa_J23022 J23022])+([http://partsregistry.org/Part:BBa_J23109 J23109]+[http://partsregistry.org/Part:BBa_J23032 J23032]). This time, B0032 was first dephosphorylated.

Dry Lab

Modeling

Association and dissociation rates have been reinvestigated (HSL with LuxR), wrong values have been removed, LuxR autoactivation has been further incorporated; we've also included diffusion of HSL.

Wiki

Small updates for the wiki: adjusting pictograms, home page, ...

Started defining a global css, intermediate result here: Team:KULeuven/Zandbak/Global

Remarks

TGS2.PNG
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