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.

Franek

Task:

• Alkaline lysis of the plasmid containing BBa_B0024
• Setting cultures with BBa_R0010 and BBa_R0080

Methods:

• PlasmidMini set by A&A Biotechnology was used. 2 test tubes with 5ml LB and Ampicyline were first inoculated with colonies containing BBa_B0024 brick on pSB1A2 plasmid. The cultures were incubated for 6h at 37C.Alkaline lysis was performed on both cultures. The pellet from 5 ml of bacteria was used.
• 3 test tubes with 5ml LB and Ampicyline were inoculated with colonies containing BBa_R0010 brick on pSB1A2 plasmid. The cultures were left for over night incubation at 37C.
• 3 test tubes with 5ml LB and Ampicyline were inoculated with colonies containing BBa_R0080 brick on pSB1A2 plasmid. The cultures were left for over night incubation at 37C.

Results:

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

Notes:

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

Jarek

Task:

• Digestion of the parts C0051 and B0032
• Electrophoretic separation of digested parts
• Isolation of DNA samples from gel
• Ligation of part C0051 to the wector with B0032

Methods:

• DNA samples were digesteg with 1 ul 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 A&A "Gel-out" kit.
• For ligation 4ul of ligase buffer and 2 ul 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 ul of plasmid solution to the bacteria. If you want to see detailed procedure go here
• Bacteria was plated on the medium containing canamycin