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 [http://partsregistry.org/Part:BBa_K177024 BBa_K177024] and [http://partsregistry.org/Part:BBa_K177025 BBa_K177025])

Franek

Tasks:

• Alkaline lysis of the plasmid containing [http://partsregistry.org/Part:BBa_B0024 BBa_B0024]

• Setting cultures with [http://partsregistry.org/Part:BBa_B0010 BBa_R0010] and [http://partsregistry.org/Part:BBa_B0080 BBa_R0080]

Methods:

• [http://aabiot.com/products/dna_purification/dna_fragments/gel_out/protocol_gel_out.pdf PlasmidMini set by A&A Biotechnology] was used. 2 test tubes with 5ml LB and ampicillin were first inoculated with colonies containing [http://partsregistry.org/Part:BBa_B0024 BBa_B0024] brick on [http://partsregistry.org/Part:pSB1A2 pSB1A2] plasmid. The cultures were incubated for 6h at 37°C. Alkaline lysis was performed on both cultures. The pellet from 5 ml of bacteria was used.

• DNA that should contain [http://partsregistry.org/Part:BBa_R0080 pAraC] on [http://partsregistry.org/Part:pSB1A2 pSB1A2] plasmid was digested with BamHI PvuI, [http://partsregistry.org/Part:BBa_R0010 placI] on [http://partsregistry.org/Part:pSB1A2 pSB1A2] was digested with IPvuI and PvuII . Digestion mix contained 5µl of extracted DNA, 2µl of Fermentas BamHI buffer , 0.3µl of each enzyme and water added to obtain 20µl total volume.

Results:

• Concentration of DNA sample was measured using NanoDrop ND-1000

Notes:

• Plates with [http://partsregistry.org/Part:BBa_I0500 BBa_I0500] transformants were empty once again. This result, together with note in [http://partsregistry.org/partsdb/get_part.cgi?part=BBa_I0500 part description] suggest that this part is damaged

Jarek

Task:

• Digestion of parts C0051 and 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 µl of PstI/BcuI (B0032) or PstI/XbaI (C0051) in 1xTango buffer for 3 hours.
• After digestion they were separated on 0,8% agarose gel.
• Isolation of sample from gel was performed with the A&A "Gel-out" kit.
• For ligation 4µl of ligase buffer and 2 µl of ligase were used.

Results:

Cloning the p53 coding sequence

Marcin

Comment:

Due to force problem with obtaining 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

Procedure:

• The protocol of transformation has not been changed, except of giving 1 µl of plasmid solution to the bacteria. If you want to see detailed procedure go here
• Bacteria was plated on the medium containing kanamycin

Construction of [http://partsregistry.org/wiki/index.php?title=Part:BBa_K177012 K177012] operon1_part2

Ania

Tasks:

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