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Team:Potsdam Bioware/Lab/Labjournal/October
From 2012.igem.org
Contents |
AID
2012-09-03
Miniprep of mVENUS construct and wildtype AID from transfected CHO cells
Investigators:
Rico
Aim:
- Miniprep of YFP-construct and wildtype AID from transfected CHO cells to proof whether it is possible to transform E.coli with the purified construct for sequencing
Method:
- Miniprep of cells
- Elution with 50 µL
Results:
CHO transfected with YFP WT-AID = 2.8 ng/µL
Antibody
2012-09-01
Glycerol stock and Mini Prep of Geneart construct, Venus, pAK100+scFv425 R H1, R H2, R L1, R L2, L1, L2
Investigator:Maria
Time: 2012-09-01 9:30 am
Materials: overnight cultures Geneart construct (4x), Venus (Tom), pAK100+scFv425 R H1, R H2, R L1, R L2, L1, L2 (Chris), Mini prep kit
Methods: according to manual
Results:
Venus: …. ng/µl
pAK100+scFv425 R H1: --- ng/µl
pAK100+scFv425 R H2: --- ng/µl<br<
pAK100+scFv425 R L1: --- ng/µl
pAK100+scFv425 R L2: --- ng/µl
pAK100+scFv425 L1: --- ng/µl
pAK100+scFv425 L2: --- ng/µl
Geneart construct 1: --- ng/µl
Geneart construct 2: --- ng/µl
Geneart construct 3: --- ng/µl
Geneart construct 4: --- ng/µl
+
Glycerole stocks of Geneart construct and pAK100+scFv425 R H1, R H2, R L1, R L2, L1, L2
Further tasks:
Virus
2012-09-02
Topic: preparing viral stocks
Investigator: Kathi
Aim: preparation of the primary virus stock
Materials:
- cell-culture --> HEK AAV 293 transfected cells on 29.08.2012 (with following plasmids: p01, p06, phelper, psb1c3)
- 80 °C freezer
- 37 °C water bath
Method:
- transfer transfected cells (incl. DMEM) to a falcon tube
- 4 times: 10 min freeze the falcon tube (-80 °C) and 15 min unfreezing (37 °C water bath)
- centrifugation (10 min, 10000 g, RT)
- supernatant = virus stock
- sore at -80 °C
Results:
- virus stock → with YFP on the surface and GOI = CFP
Further tasks
- infection
