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Team:Potsdam Bioware/Lab/Labjournal/October
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<b>Topic: </b>cell-culture <br> | <b>Topic: </b>cell-culture <br> | ||
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| - | * | + | * transfection of new scFv RAGE construct |
| - | * | + | * transfection of TAL-AID from cooperation with Freiburg iGEM Team in CHO cells (co-transfection with clone 4) |
| - | * | + | * purification of Nanobody from supernatant of Cre recombinase transfected cells with magnetic beads |
| - | * | + | * Western Blot of purified Nanobody |
==Virus== | ==Virus== | ||
Revision as of 13:59, 25 October 2012
Contents |
AID
2012-10-10
Inoculation of plasmid samples of the 48h retransformation plates (after FACS)
Investigators: Tom
Time: 2012-10-10
Materials:
- LB medium
- Amp stock solution
- plate with cultures:
EGFR-C-AID without NES, with NLS+Kozak sequence+eGFP
(after FACS selection)
Method:
Inoculation of:
5 cultures per plate in 5 ml LB medium + 5µL Amp.
(--> 5 cultures)
Further tasks:
- Miniprep
Antibody
2012-10-12
cell culture
Investigator:Kerstin
Topic: cell-culture
- passaging of CHO w Zeocin
- passaging of HT1080
- passaging of HEK AAV 293
- passaging of HeLa
2012-10-15
cell culture
Investigator:Kerstin
Topic: cell-culture
- passaging of CHO w Zeocin
- passaging of HT1080
- passaging of HEK AAV 293
- passaging of HeLa
2012-10-17
cell culture
Investigator:Kerstin
Topic: cell-culture
- passaging of CHO w Zeocin
- passaging of HT1080
- passaging of HEK AAV 293
- passaging of HeLa
- passaging of stably transfected Clones 4.1, 4.2, 4.3, 4,4, 29.1, 29.2, 29.3 with 550µg/ml Hygromycin
2012-10-19
cell culture
Investigator:Kerstin
Topic: cell-culture
- passaging of CHO w Zeocin
- passaging of HT1080
- passaging of HEK AAV 293
- passaging of HeLa
2012-10-22
cell culture
Investigator:Kerstin
Topic: cell-culture
- passaging of CHO w Zeocin
- passaging of HT1080
- passaging of HEK AAV 293
- passaging of HeLa
- transfection of stably transfected clone 29.1 with Cre Recombinase
- seeding of HT1080 in Ibidi Dish for infection with virus
2012-10-23
cell culture
Investigator:Kerstin
Topic: cell-culture
- seeding of CHO and HeLa in Ibidi Dishes for transfection with new Nanobody RAGE construct
- infection with Virus (CFP on surface and YFP as GOI, AAV with Sortase motif)
2012-10-24
cell culture
Investigator:Kerstin
Topic: cell-culture
- Transfection of new Nanobody RAGE construct in CHO and HeLa
- seeding of CHO and HeLa in Ibidi Dishes for transfection with new scFv RAGE construct
- passaging of CHO w Zeocin
- passaging of HT1080
- passaging of HEK AAV 293
- passaging of HeLa
2012-10-25
cell culture
Investigator:Kerstin
Topic: cell-culture
- transfection of new scFv RAGE construct
- transfection of TAL-AID from cooperation with Freiburg iGEM Team in CHO cells (co-transfection with clone 4)
- purification of Nanobody from supernatant of Cre recombinase transfected cells with magnetic beads
- Western Blot of purified Nanobody
Virus
XXX
XXX
Investigator: XXX
Aim: XXX
Materials:
XXX
Method:
XXX
Results:
XXX
Further tasks
XXX
