Revision as of 13:16, 7 October 2009

Meetings Calendar

Formal Lab Session - 15th September 2009

Stochastic Switch Team

Today we set up another restriction disest of the KinA pMKRQ synthesised brick using NheI and EcoRI we ran this on an 0.8% gel however we think that this may be too dilute and that I why we are not seeing the two bands ogf the digest which are a similar size.

We also set up another PCR of the ara fragment as a back up plan for ligation, in case the last ligation did not work as well as we had planned.

We also mini prepped and digested the sac transformations again, and we found that of the 6 samples we had, a few of them had worked!

Just to confirm however we set up 12 more miniprep cultures again for both sac and ara.

Yesterday we did midipreps of Goksels successful sspb plasmid: this was cut with XbaI and PstI ready for ligation into the ara brick (once it's ligated!).

Metal Sensing Team

Introduction

Yesterday's lab session saw the Metal Sensing team ligate the cadA promoter region with the linearised BBa_J33206 BioBrick (BioBrick containing arsR gene and binding site but with promoter absent). The team also attempted to ligate cotC and linearised pMUTIN4. Both of these ligated products were used to transform DH5-alpha E. coli cells.

Today's work will involve the team looking for transformant colonies on the plates and processing them based on observations. If there are colonies, 12 will be picked and used to inoculate 12 tubes of 5ml LB (+ antibiotic) for mini-preps. If unsuccessful, another attempt at ligation and then transformation will follow. The team will also carry out digests of the 5 midi-preps we had done earlier to prove that they have the correct DNA.

Practical Outline

Observations

Procedure

Preparation for cotC transformant mini-preps

Digesting midi-prep samples

Ligation of cadA promoter and arsR BioBrick

Transformation of E. coli with ligated product

20 – 21 June 2009 - Europe workshop (London)
23 – 24 June 2009 - UK iGEM meetup (Edinburgh)
23 October Practice Presentation (Newcastle)
23 October T-shirts are ready
27 October Practice Presentation (Sunderland)
27 October Poster is ready
30 October – 2 November 2009 - Jamboree (Boston)

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[http://www.facebook.com/home.php#/group.php?gid=131709337641 Newcastle on Facebook]
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@@ Line 19: / Line 19: @@
 ==<u>Metal Sensing Team</u>==
 ===Introduction===
+Yesterday's lab session saw the Metal Sensing team ligate the ''cadA'' promoter region with the linearised ''BBa_J33206'' BioBrick (BioBrick containing ''arsR'' gene and binding site but with promoter absent). The team also attempted to ligate ''cotC'' and linearised ''pMUTIN4''. Both of these ligated products were used to transform ''DH5-alpha'' ''E. coli'' cells.
+'''Today's work will involve the team looking for transformant colonies on the plates and processing them based on observations.''' If there are colonies, 12 will be picked and used to inoculate 12 tubes of 5ml LB (+ antibiotic) for mini-preps. If unsuccessful, another attempt at ligation and then transformation will follow. '''The team will also carry out digests of the 5 midi-preps we had done earlier to prove that they have the correct DNA'''.
 ===Practical Outline===
 ===Observations===

Team:Newcastle/Labwork/15 September 2009

From 2009.igem.org