EricChu:LabNotesMDA/2014-2-18
Jump to navigation
Jump to search
Adding N10(unmodified) into KOH as blocking agent in v7 device
- 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.
File:02192014N10qPCRAlu.jpg File:02192014N10qPCRMito.jpg
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.
- 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