User:Anthony Salvagno/Notebook/Research/2011/03/04/Prepping Unzipping With pALS: Difference between revisions
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I only have SappBR so I will need to ligate the adapter to here first and then ligate this to the digested pALS. | 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=== | ===SappBR-Adapter Ligation=== | ||
{{ShowGoogleExcel|id=0Agbdciapt4QZdHZpSjF5UThUek5JN3NtR1RtUkFoenc|width=500|height=300}} | |||
===ApBR-DpALS Ligation=== | ===ApBR-DpALS Ligation=== | ||
There is a good chance this doesn't happen today. | There is a good chance this doesn't happen today. | ||
Revision as of 11:16, 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 }}
ApBR-DpALS Ligation
There is a good chance this doesn't happen today.
