User:Anthony Salvagno/Notebook/Research/2011/03/04/Prepping Unzipping With pALS: Difference between revisions
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**151.1ng/ul --> ~57nM -- ~4.5ng | **151.1ng/ul --> ~57nM -- ~4.5ng | ||
===ApBR-DpALS Ligation=== | |||
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[[Category:Ant Digestion]] | [[Category:Ant Digestion]] | ||
[[Category:Ant Ligation]] | [[Category:Ant Ligation]] | ||
[[Category:Ant Anneal]] | [[Category:Ant Anneal]] |
Latest revision as of 14:02, 4 March 2011
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PCR
I cleaned up the PCR reaction from Thursday and the nanodrop revealed:
- 227ng/ul --> 85nM in 30ul (total of 6.8ug of DNA)
BstXI Digestion
{{#widget:Google Spreadsheet |key=0Agbdciapt4QZdFRQdlV3cGV5eHgzTEE2VnZGRGlua3c |width=500 |height=300 }}
Adapter Annealing
Since I got a new bottom oligo with 5' biotinylation I will have to anneal a new adapter duplex. {{#widget:Google Spreadsheet |key=0Agbdciapt4QZdGVoNElGZ0piZ0FzR0hGOFdBWkhZdGc |width=500 |height=300 }}
Ligation
I only have SappBR so I will need to ligate the adapter to here first and then ligate this to the digested pALS.
SappBR-Adapter Ligation
{{#widget:Google Spreadsheet |key=0Agbdciapt4QZdHZpSjF5UThUek5JN3NtR1RtUkFoenc |width=500 |height=300 }}
After purification:
- ApBR:
- 89ng/ul --> ~31nM -- ~2.7ug
- DpALS:
- 151.1ng/ul --> ~57nM -- ~4.5ng
ApBR-DpALS Ligation
{{#widget:Google Spreadsheet |key=0Agbdciapt4QZdDcyZ2JEanhRS3FRTUNqc2luMUM1Unc |width=500 |height=300 }}