EricChu:LabNotesMDA/2014-2-18: Difference between revisions
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* Final N10 concentration was 10uM. | * Final N10 concentration was 10uM. | ||
[[File:02192014N10qPCRAlu.jpg]] | * qPCR of Alu after sample collection: | ||
[[File:02192014N10qPCRMito.jpg]] | [[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. | * 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:
- 8ul 5M KOH
- 5ul 2M DTT
- 18.75ul 8% Tween20
- 10ul 100uM N10
- 58.25ul water
- Final N10 concentration was 10uM.
- qPCR of Alu after sample collection:
- qPCR of Mito after sample collection:
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