Daniel:Notebook/PosSequencing/2017-5-17: Difference between revisions
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>Djacobse (Created page with "=Bead Binding Test 3= Back to Calendar Trying to test the bead binding and denaturation on the beads again. This time we'll use high pH as ...") |
>Djacobse |
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[[Daniel:Notebook/PosSequencing|Back to Calendar]] | [[Daniel:Notebook/PosSequencing|Back to Calendar]] | ||
Trying to test the bead binding and denaturation on the beads again. This time we'll use high pH as the denaturant. | [[Media:Kohler_Avidin-Streptavidin_Bonds_Molecules_2014.pdf|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 [https://biology.stackexchange.com/questions/29925/why-does-high-ph-result-in-the-denaturation-of-dna 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. | |||
==Protocol== | ==Protocol== |
Revision as of 15:39, 17 May 2017
Bead Binding Test 3
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.