Team:Heidelberg/Notebook measure/NotebookDP
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__NOTOC__ | __NOTOC__ | ||
- | =="Dual plasmid"== | + | ==""Dual plasmid""== |
== Contents == | == Contents == | ||
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|style="text-align:center"| [[Team:Heidelberg/Notebook_measure/NotebookDP#8-27-2009|8-27-2009]] | |style="text-align:center"| [[Team:Heidelberg/Notebook_measure/NotebookDP#8-27-2009|8-27-2009]] | ||
- | |style="text-align:center"| - | + | |style="text-align:center"| [[Team:Heidelberg/Notebook_measure/NotebookDP#8-28-2009|8-28-2009]] |
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|style="text-align:center"| - | |style="text-align:center"| - | ||
+ | |style="text-align:center"| [[Team:Heidelberg/Notebook_measure/NotebookDP#9-01-2009|9-01-2009]] | ||
+ | |style="text-align:center"| [[Team:Heidelberg/Notebook_measure/NotebookDP#9-02-2009|9-02-2009]] | ||
+ | |style="text-align:center"| [[Team:Heidelberg/Notebook_measure/NotebookDP#9-03-2009|9-03-2009]] | ||
+ | |style="text-align:center"| [[Team:Heidelberg/Notebook_measure/NotebookDP#9-04-2009|9-04-2009]] | ||
+ | |style="text-align:center"| [[Team:Heidelberg/Notebook_measure/NotebookDP#9-05-2009|9-05-2009]] | ||
+ | |style="text-align:center"| [[Team:Heidelberg/Notebook_measure/NotebookDP#9-06-2009|9-06-2009]] | ||
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|style="text-align:center"| 37 | |style="text-align:center"| 37 | ||
- | |style="text-align:center"| - | + | |style="text-align:center"| [[Team:Heidelberg/Notebook_measure/NotebookRT#9-07-2009|9-07-2009]] |
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|style="text-align:center"| [[Team:Heidelberg/Notebook_measure/NotebookRT#9-10-2009|9-10-2009]] | |style="text-align:center"| [[Team:Heidelberg/Notebook_measure/NotebookRT#9-10-2009|9-10-2009]] | ||
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** redo BclI digest later | ** redo BclI digest later | ||
- | ==9- | + | ==9-01-09== |
* Transformation of PCR_mCherry_p7 | * Transformation of PCR_mCherry_p7 | ||
** should have been in unmethylating cells, but wasn't--> no test-digest with BclI possible | ** should have been in unmethylating cells, but wasn't--> no test-digest with BclI possible | ||
** control result via PCR--> Product can be used for further cloning | ** control result via PCR--> Product can be used for further cloning | ||
- | ==9- | + | ==9-02-09== |
* picked colonies from transformation of PCR_mCherry_p7 | * picked colonies from transformation of PCR_mCherry_p7 | ||
* PCR of minipreps from ligation PCR_mCherry_p7 with O.172 and O.173 to amplify CMV_mCherry with SphI restriction sites on both ends | * PCR of minipreps from ligation PCR_mCherry_p7 with O.172 and O.173 to amplify CMV_mCherry with SphI restriction sites on both ends | ||
- | ==9- | + | ==9-03-09== |
* redid Bcl1/Nhe1 digest of p7: at least 9 clear bands visible on 1% argarose gel--> has to be redone again | * redid Bcl1/Nhe1 digest of p7: at least 9 clear bands visible on 1% argarose gel--> has to be redone again | ||
* PCR purification of PCR_mCherry_CMV ( 8 samples from ligation) | * PCR purification of PCR_mCherry_CMV ( 8 samples from ligation) | ||
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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- | + | ==9-04-2009== |
* 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- | + | ==9-05-2009== |
* 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- | + | ==9-06-2009== |
* 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- | + | ==9-07-2009== |
* 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 |
Revision as of 18:53, 21 October 2009
""Dual plasmid""Contents
8-27-09
8-28-09
9-01-09
9-02-09
9-03-09
9-04-2009
--> ligation PCR_mCherry_CMV and p31 can not have worked either
9-05-2009
--> Repeat from the beginning
9-06-2009
9-07-2009
9-10-2009
|