Team:Heidelberg/Notebook measure/NotebookDP
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__NOTOC__ | __NOTOC__ | ||
+ | =="Dual plasmid"== | ||
+ | |||
+ | == Contents == | ||
+ | |||
+ | {| class="wikitable centered" border="2" rules="rows" width="900px" style="border-color:white;" | ||
+ | |- | ||
+ | ! Week !! colspan="7" |Days | ||
+ | |- | ||
+ | |style="text-align:center"| 35 | ||
+ | |style="text-align:center"| - | ||
+ | |style="text-align:center"| - | ||
+ | |style="text-align:center"| - | ||
+ | |style="text-align:center"| [[Team:Heidelberg/Notebook_measure/NotebookDP#8-27-2009|8-27-2009]] | ||
+ | |style="text-align:center"| - | ||
+ | |style="text-align:center"| - | ||
+ | |style="text-align:center"| - | ||
+ | |- | ||
+ | |style="text-align:center"| 36 | ||
+ | |style="text-align:center"| [[Team:Heidelberg/Notebook_measure/NotebookRT#8-31-2009|8-31-2009]] | ||
+ | |style="text-align:center"| - | ||
+ | |style="text-align:center"| [[Team:Heidelberg/Notebook_measure/NotebookRT#9-02-2009|9-02-2009]] | ||
+ | |style="text-align:center"| [[Team:Heidelberg/Notebook_measure/NotebookRT#9-03-2009|9-03-2009]] | ||
+ | |style="text-align:center"| [[Team:Heidelberg/Notebook_measure/NotebookRT#9-04-2009|9-04-2009]] | ||
+ | |style="text-align:center"| - | ||
+ | |style="text-align:center"| - | ||
+ | |- | ||
+ | |style="text-align:center"| 37 | ||
+ | |style="text-align:center"| - | ||
+ | |style="text-align:center"| - | ||
+ | |style="text-align:center"| - | ||
+ | |style="text-align:center"| [[Team:Heidelberg/Notebook_measure/NotebookRT#9-10-2009|9-10-2009]] | ||
+ | |style="text-align:center"| - | ||
+ | |style="text-align:center"| - | ||
+ | |style="text-align:center"| - | ||
+ | |- | ||
+ | |style="text-align:center"| 39 | ||
+ | |style="text-align:center"| [[Team:Heidelberg/Notebook_measure/NotebookRT#9-21-2009|9-21-2009]] | ||
+ | |style="text-align:center"| - | ||
+ | |style="text-align:center"| - | ||
+ | |style="text-align:center"| [[Team:Heidelberg/Notebook_measure/NotebookRT#9-24-2009|9-24-2009]] | ||
+ | |style="text-align:center"| [[Team:Heidelberg/Notebook_measure/NotebookRT#9-25-2009|9-25-2009]] | ||
+ | |style="text-align:center"| - | ||
+ | |style="text-align:center"| - | ||
+ | |- | ||
+ | |style="text-align:center"| 40 | ||
+ | |style="text-align:center"| - | ||
+ | |style="text-align:center"| - | ||
+ | |style="text-align:center"| [[Team:Heidelberg/Notebook_measure/NotebookRT#9-30-2009|9-30-2009]] | ||
+ | |style="text-align:center"| - | ||
+ | |style="text-align:center"| [[Team:Heidelberg/Notebook_measure/NotebookRT#10-02-2009|10-02-2009]] | ||
+ | |style="text-align:center"| - | ||
+ | |style="text-align:center"| - | ||
+ | |- | ||
+ | |} | ||
+ | |||
+ | |||
==8-27-09== | ==8-27-09== | ||
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** BclI can not be heat-inactivated! | ** BclI can not be heat-inactivated! | ||
- | == | + | ==8-28-09== |
* restriction digest of p7 and PCR_mCherry w/ BclI for 2 hours at 50°C | * restriction digest of p7 and PCR_mCherry w/ BclI for 2 hours at 50°C | ||
** looked at result on agarose gel: 3 clear bands, where only two are suppossed to be | ** looked at result on agarose gel: 3 clear bands, where only two are suppossed to be | ||
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* transfection of the ligation PCR_mCherry in p7 from 8-28-09 in HeLa cells according to effectene protocol | * transfection of the ligation PCR_mCherry in p7 from 8-28-09 in HeLa cells according to effectene protocol | ||
- | == | + | ==9-4-2009== |
* transfection of the ligation PCR_mCherry in p7 from 8-28-09 did not work, but positive control worked: ligation not successful | * transfection of the ligation PCR_mCherry in p7 from 8-28-09 did not work, but positive control worked: ligation not successful | ||
--> ligation PCR_mCherry_CMV and p31 can not have worked either | --> ligation PCR_mCherry_CMV and p31 can not have worked either | ||
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** sent for sequencing | ** sent for sequencing | ||
- | == | + | ==9-5-2009== |
* gel purification of MfeI_Jet_NheI | * gel purification of MfeI_Jet_NheI | ||
* restriction digest of MfeI_Jet__NheI and p6 with MfeI and NheI | * restriction digest of MfeI_Jet__NheI and p6 with MfeI and NheI | ||
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* picked 8 colonies of the latest p7::mcherry transformation | * picked 8 colonies of the latest p7::mcherry transformation | ||
- | == | + | ==9-6-2009== |
* Miniprep of p7::mcherry | * Miniprep of p7::mcherry | ||
** extraction of CMV::mcherry from p7::mcherry by PCR with O.172 and O.173 | ** extraction of CMV::mcherry from p7::mcherry by PCR with O.172 and O.173 | ||
* extraction of Jet with MfeI and NheI restriction sites from p31 by PCR (O.203 and O.71) | * extraction of Jet with MfeI and NheI restriction sites from p31 by PCR (O.203 and O.71) | ||
- | == | + | ==9-7-2009== |
* PCR-purification of PCR_mCherry_CMV and PCR_Mfe_Jet_Nhe | * PCR-purification of PCR_mCherry_CMV and PCR_Mfe_Jet_Nhe | ||
* restriction digest of p31 and PCR_mCherry_CMV with Sph1 | * restriction digest of p31 and PCR_mCherry_CMV with Sph1 | ||
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* preparative gelelectrophoresis of the restriction digests | * preparative gelelectrophoresis of the restriction digests | ||
- | == | + | ==9-10-2009== |
* ligated mCherry (digested w/ BclI, AvrII) with p7 (BclI, NheI) and p7JeT (BclI, NheI) overnight | * ligated mCherry (digested w/ BclI, AvrII) with p7 (BclI, NheI) and p7JeT (BclI, NheI) overnight |
Revision as of 18:46, 21 October 2009
"Dual plasmid"Contents
8-27-09
8-28-09
9-1-09
9-2-09
9-3-09
9-4-2009
--> ligation PCR_mCherry_CMV and p31 can not have worked either
9-5-2009
--> Repeat from the beginning
9-6-2009
9-7-2009
9-10-2009
|