User:Wilfred J. Poppinga/Notebook/cAMP compartmentalization/2010/07/02: Difference between revisions

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{|{{table}} width="800"
{|{{table}} width="800"
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|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> Project name</span>
|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;">qPCR GAPDH, Silverstaining</span>
|style="background-color: #F2F2F2" align="center"|<html><img src="/images/9/94/Report.png" border="0" /></html> [[{{#sub:{{FULLPAGENAME}}|0|-11}}|Main project page]]<br />{{#if:{{#lnpreventry:{{FULLPAGENAME}}}}|<html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>[[{{#lnpreventry:{{FULLPAGENAME}}}}{{!}}Previous entry]]<html>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</html>}}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]]<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>}}
|style="background-color: #F2F2F2" align="center"|<html><img src="/images/9/94/Report.png" border="0" /></html> [[{{#sub:{{FULLPAGENAME}}|0|-11}}|Main project page]]<br />{{#if:{{#lnpreventry:{{FULLPAGENAME}}}}|<html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>[[{{#lnpreventry:{{FULLPAGENAME}}}}{{!}}Previous entry]]<html>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</html>}}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]]<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>}}
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===Method===
===Method===
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|+'''6-wells'''
|+'''48-wells plate'''
!#
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|+'''24-wells'''
!#
!<u>1</u>
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!<u>5</u>
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!07
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|(-) GAPDH
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*After doing the Coomassie staining a smear could be vaguely distuingished around 250 kDa
*After doing the Coomassie staining a smear could be vaguely distuingished around 250 kDa
** But during the meeting today results were presented showing that the Enspire 2300 has a malfunction leading to wrong protein concentrations
** But during the meeting today results were presented showing that the Enspire 2300 has a malfunction leading to wrong protein concentrations
*qPCR results for AKAP12
[[Image:06072010_qPCR_AKAP12.png|500px]]


==Discussion==
==Discussion==
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==Attachment==
==Attachment==
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<html>
<a href="http://openwetware.org/wiki/Image:HTERT_Donor_12_ht31_160310.xls">
<a href="http://openwetware.org/wiki/Image:02072010_qPCR_GAPDH.xls">
   <img src="http://openwetware.org/images/f/f8/Owwnotebook_icon.png" alt="" width="50" height="50">
   <img src="http://openwetware.org/images/f/f8/Owwnotebook_icon.png" alt="" width="50" height="50">
</a>
</a>
</html> [[Media:FILE.xls| Excel TITLE]]
</html> [[Media:02072010_qPCR_GAPDH.xls|qPCR for GAPDH]]
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__NOTOC__
__NOTOC__

Revision as of 05:28, 29 July 2010

<html><style type="text/css"> .todo { color: red } .done { color: green} </style></html>

qPCR GAPDH, Silverstaining <html><img src="/images/9/94/Report.png" border="0" /></html> Main project page
<html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>Previous entry<html>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</html>Next entry<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>

Summary

GAPDH

As internal control also GAPDH need to be measured,

  • 20 uL reaction volume, as given by the machine
  • Human GAPDH primer (Applied biosystems)
  • Taqman Gene expression master mix

Silver staining

  • Repeat of yesterday on same gel, fresh 0.1% Silver nitrate was prepared

Materials & Methods

Materials

Method

48-wells plate
# 01 02 03 04 05 06 07 08
A S1 S1 S1 (-) GAPDH (-) GAPDH (-) GAPDH
B S2 S2 S2
C S3 S3 S3
D C1 C1 C1
E C2 C2 C2
F C3 C3 C3

Notes

  • 48-wells plate was not spinned down before PCR

Results

  • After doing the Coomassie staining a smear could be vaguely distuingished around 250 kDa
    • But during the meeting today results were presented showing that the Enspire 2300 has a malfunction leading to wrong protein concentrations
  • qPCR results for AKAP12

Discussion

  • DONE Ow..

Related

Related entries

Same actions

Related topics

Attachment

<html> <a href="http://openwetware.org/wiki/Image:02072010_qPCR_GAPDH.xls"> 

  <img src="http://openwetware.org/images/f/f8/Owwnotebook_icon.png" alt="" width="50" height="50">

</a> </html> qPCR for GAPDH


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