User:Anthony Salvagno/Notebook/Research/2011/03/04/Prepping Unzipping With pALS

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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)

Adapter Annealing

Since I got a new bottom oligo with 5' biotinylation I will have to anneal a new adapter duplex.

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

After purification:

• ApBR:
  • 89ng/ul --> ~31nM -- ~2.7ug
• DpALS:
  • 151.1ng/ul --> ~57nM -- ~4.5ng