Team:Potsdam Bioware/Lab/Labjournal/October

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==AID==
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===<p style="background-color: rgb(240, 20, 70);">2012-09-03</p>===
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<p style="background-color: rgb(238, 221, 130); font-weight: bold;">Miniprep of mVENUS construct and wildtype AID from transfected CHO cells</p>
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<Br>
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<b>Investigators:</b><Br>
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 +
Rico
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 +
<Br>
 +
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<b>Aim:</b><Br>
 +
 +
*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
 +
 +
<b>Method:</b><Br>
 +
 +
*Miniprep of cells
 +
 +
*Elution with 50 µL
 +
 +
<Br>
 +
 +
<b>Results:</b><Br>
 +
 +
CHO transfected with YFP WT-AID = 2.8 ng/µL<Br>
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<Br>
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==Antibody==
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===<p style="background-color: rgb(240, 20, 70);">2012-09-01</p>===
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<p style="background-color: rgb(238, 221, 130); font-weight: bold;">Glycerol stock and Mini Prep of Geneart construct, Venus, pAK100+scFv425 R H1, R H2, R L1, R L2, L1, L2 </p>
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<br>
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<b>Investigator:</b>Maria<br>
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<br>
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<b>Time: </b>2012-09-01 9:30 am<br>
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<br>
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<b>Materials: </b>overnight cultures Geneart construct (4x), Venus (Tom), pAK100+scFv425 R H1, R H2, R L1, R L2, L1, L2 (Chris), Mini prep kit <br>
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<br>
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<b>Methods: </b>according to manual <br>
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<br>
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<b>Results: </b>
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Venus: …. ng/µl<br>
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pAK100+scFv425 R H1: --- ng/µl<br>
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pAK100+scFv425 R H2: --- ng/µl<br<
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pAK100+scFv425 R L1: --- ng/µl<br>
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pAK100+scFv425 R L2: --- ng/µl<br>
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pAK100+scFv425 L1: --- ng/µl<br>
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pAK100+scFv425 L2: --- ng/µl<br>
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Geneart construct 1: --- ng/µl<br>
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Geneart construct 2: --- ng/µl<br>
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Geneart construct 3: --- ng/µl<br>
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Geneart construct 4: --- ng/µl<br>
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+ <br>
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Glycerole stocks of Geneart construct and pAK100+scFv425 R H1, R H2, R L1, R L2, L1, L2
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<br>
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<b>Further tasks: </b><br>
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<br>
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==Virus==
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===<p style="background-color: rgb(240, 20, 70);"> 2012-09-02</p>===
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<p style="background-color: rgb(238, 221, 130); font-weight: bold;"> Topic: preparing viral stocks</p>
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<b>Investigator: </b> Kathi <br>
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<b> Aim: </b> preparation of the primary virus stock<br>
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<b>Materials:</b><br>
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* cell-culture --> HEK AAV 293 transfected cells on 29.08.2012 (with following plasmids: p01, p06, phelper, psb1c3)
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* 80 °C freezer
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* 37 °C water bath <br>
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<b>Method:</b><br>
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* transfer transfected cells (incl. DMEM) to a falcon tube
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* 4 times: 10 min freeze the falcon tube (-80 °C) and 15 min unfreezing (37 °C water bath)
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* centrifugation (10 min, 10000 '''g''', RT)
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* supernatant = virus stock
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* sore at -80 °C <br>
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<b>Results:</b><br>
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* virus stock → with YFP on the surface and GOI = CFP <br>
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<b>Further tasks</b>
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* infection

Revision as of 09:24, 22 October 2012


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

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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
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