Team:Warsaw/Calendar-Main/10 July 2009

Team meeting


 * 1) presentations of work did by both groups during last week (given by Ania and Kuba)
 * 2) presentation of methods used by teams starting in previous editions of iGEM to kill bacteria :) (Kamil)
 * 3) presentation of different methods of targeting drugs to cancer cells (Marcin)

We've been also talking about some organization issues and we've decided to move our meetings - now they will take place every Thursady at 17:00, one week at the Faculty of Biology Building, another week in the room E of the lab on Pawinskiego Street.

Creating devices to test promoters in E. coli strains (devices BBa_K177024 and BBa_K177025)
Franek

Tasks:

Methods:
 * Alkaline lysis of the plasmid containing BBa_B0024 terminator

Results:
 * PlasmidMini set by A&A Biotechnology was used. 2 test tubes with 5ml LB and ampicillin were first inoculated with colonies containing BBa_B0024 brick on pSB1A2 plasmid. The cultures were incubated for 6h at 37&deg;C. Alkaline lysis was performed on both cultures. The pellet from 5 ml of bacteria was used.

Notes:
 * Concentration of DNA sample was measured using NanoDrop ND-1000


 * Plates with BBa_I0500 transformants were empty once again. This result, together with note in part description suggest that this part is damaged

Making of RBS-cI part
Jarek Task:  Digestion of parts  BBa_C0051 and  BBa_B0032  Electrophoretic separation of digested parts Isolation of DNA samples from gel Ligation of part C0051 to the vector with B0032  Methods:  DNA samples were digesteg with 1 &micro;l of PstI/BcuI (B0032) or PstI/XbaI (C0051) in 1xTango buffer for 3 hours.</li> After digestion they were separated on 0,8% agarose gel.</li> Isolation of sample from gel was performed with the A&A "Gel-out" kit</a>.</li> For ligation 4&micro;l of ligase buffer and 2 &micro;l of ligase were used. </ul> Results:  </ul>

Cloning of p53 coding sequence Marcin Comment:  Due to problem with low quality of PCR product it was necessary to transform bacteria with plasmid containing p53 sequence and to perform all procedures once more time. Tasks:  Transformation of chemocompetent strain of E. coli by plasmid containing p53 sequence</li> </ul> Procedure:  The protocol of transformation has not been changed, except of giving 1 &micro;l of plasmid solution to the bacteria. If you want to see detailed procedure go here</a> </li> Bacteria was plated on the medium containing kanamycin</li></ul>

Construction of K177012 operon1_part2
Ania

Tasks:


 * Competent cells were transformed with LacI C0012 taken out of the distribution 2009 Kit Plate 1 well 20.

Isolation of BioBricks from 2008 and 2009 Kit Plates
Monika

Tasks:


 * Isolate plasmids containing the biobricks - continuation


 * 1) GFP coding device switched on by IPTG - BBa_I763004  from 2008 Kit Plate 1017 well G5
 * 2) promoter lambda (cI regulated) with RFP reporter BBa_I763007  from 2009 Kit Plate 1 well 15J
 * 3) GFP generator - BBa_E0840 from 2009 Kit Plate 1 well 12O
 * 4) GFP generator - BBa_J07037 from 2009 Kit Plate 1 well 12O

Results from 9 July 2009
 * all transformations exept GFP coding device switched on by IPTG (BBa_I763004) were successful

Methods:
 * Planting on LB medium supplemented with apropriate antibiotic, 6h incubation in 37&deg;C
 * Planting on LB-agar medium supplemented with apropriate antibiotic
 * Alkaline lysis (prodedure described here)

Results
 * Concentration of DNA sample was measured using NanoDrop ND-1000

Comment
 * Isolations where DNA concentration is under 30ng/&micro;l must be repeated