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

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qPCR GAPDH, Silverstaining Main project page
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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..

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Attachment

qPCR for GAPDH


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