Daniel:Notebook/PosSequencing/2017-5-17: Difference between revisions

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*8 ul 5 M KOH.
*8 ul 5 M KOH.
*10 ul 1 M DTT.
*10 ul 1 M DTT.
*2 ul 0.5 M EDTA.
*2 ul 0.5 M EDTA
 
'''NS buffer'''
 
*3 mL H20
*3 mL 2M Tris-HCl
*4 mL 1N HCl


==Protocol==
==Protocol==

Revision as of 15:56, 17 May 2017

Bead Binding Test 3

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Avidin-Biotin Bonds Paper

Trying to test the bead binding and denaturation on the beads again. This time we'll use high pH as the denaturant. From the paper above it says the streptavidin bond is stable to about pH 11, and according to this website DNA denaturation occurs at pH>10. I will use the alkaline lysis solution from Andrew's notes

ALS Buffer

  • 80 ul Nuclease free H2O.
  • 8 ul 5 M KOH.
  • 10 ul 1 M DTT.
  • 2 ul 0.5 M EDTA

NS buffer

  • 3 mL H20
  • 3 mL 2M Tris-HCl
  • 4 mL 1N HCl

Protocol