EricChu:LabNotesMDA/2014-2-18: Difference between revisions

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# 58.25ul water
# 58.25ul water
* Final N10 concentration was 10uM.
* Final N10 concentration was 10uM.
* qPCR of Alu after sample collection:
[[File:02192014N10qPCRAlu.jpg|1000px]]
* qPCR of Mito after sample collection:
[[File:02192014N10qPCRMito.jpg|1000px]]


==Adding N10(unmodified) into KOH as blocking agent in Realtime MDA==
==Adding N10(unmodified) into KOH as blocking agent in Realtime MDA==
* Beside the above condition (10uM N10), 1uM and 50uM were also tested on 100pg and 10pg genomic DNA.
* Beside the above condition (10uM N10), 1uM and 50uM were also tested on 100pg and 10pg genomic DNA.
 
* Result: N10 seemed to be inhibiting the MDA reaction. Higher concentration N10 reduced the MDA time. All 6 conditions did not amplify by Alu nor Mito primers.
[[File:02182014realtimeMDAN10.jpg]]
[[File:02182014realtimeMDAN10.jpg]]
[[File:02182014qPCRrealtimeMDAN10.jpg|1000px]]

Latest revision as of 00:10, 1 March 2014

Adding N10(unmodified) into KOH as blocking agent in v7 device[edit]

  • This experiment was done following Hosuk's Jun28 protocol (PGP1#6). The cell was pumped in ALS at 1sec interval for 10mins.
  • ALS recipe:
  1. 8ul 5M KOH
  2. 5ul 2M DTT
  3. 18.75ul 8% Tween20
  4. 10ul 100uM N10
  5. 58.25ul water
  • Final N10 concentration was 10uM.
  • qPCR of Alu after sample collection:

File:02192014N10qPCRAlu.jpg

  • qPCR of Mito after sample collection:

File:02192014N10qPCRMito.jpg

Adding N10(unmodified) into KOH as blocking agent in Realtime MDA[edit]

  • Beside the above condition (10uM N10), 1uM and 50uM were also tested on 100pg and 10pg genomic DNA.
  • Result: N10 seemed to be inhibiting the MDA reaction. Higher concentration N10 reduced the MDA time. All 6 conditions did not amplify by Alu nor Mito primers.

File:02182014realtimeMDAN10.jpg File:02182014qPCRrealtimeMDAN10.jpg