Daniel:Notebook/PosSequencing/2017-5-17: Difference between revisions
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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 [[Arichard:Reagents/als|Andrew's notes]] | 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 [[Arichard:Reagents/als|Andrew's notes]] | ||
''' | '''Alkaline Lysis Solution''' | ||
*80 ul Nuclease free H2O. | *80 ul Nuclease free H2O. | ||
Line 14: | Line 14: | ||
*2 ul 0.5 M EDTA | *2 ul 0.5 M EDTA | ||
''' | '''Neutralization Solution''' | ||
*3 mL H20 | *3 mL H20 |
Revision as of 15:57, 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
Alkaline Lysis Solution
- 80 ul Nuclease free H2O.
- 8 ul 5 M KOH.
- 10 ul 1 M DTT.
- 2 ul 0.5 M EDTA
Neutralization Solution
- 3 mL H20
- 3 mL 2M Tris-HCl
- 4 mL 1N HCl