User:Kathryn Muratore/Notebook/AU CHEM-570 lab prep/2011/06/20

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 * style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]] AU CHEM-570 Lab Prep
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 * style="background-color: #F2F2F2" align="center"|  |Main project page


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Bench work

 * 1) PCR cleanup
 * 2) * use | Promega Wizard SV Gel and PCR Cleanup kit.
 * 3) 70μL of membrane binding solution added directly to  BSA PCR tube, for a final volume of 140μL (some of the extra volume is from the wax, but this is still more than expected assuming that I used 15μL from this tube  last week).
 * 4) followed manufacturer protocol, using the vacuum method, and eluted with 50μL H2O
 * 5) *&rarr; store @ -20°C
 * 6) NheI digest
 * 7) 15μL of cleaned-up BSA PCR from step 1 + 5μL NEB buffer 2 + 24.5μL H2O + 0.5μL 100X BSA + 5μL NheI
 * 8) *&rarr; 2h@37°C
 * 9) *&rarr; 20'@65°C
 * 10) *&rarr; store @ -20°C


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