User:Kathryn Muratore/Notebook/AU CHEM-570 lab prep/2011/06/14

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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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Plan
(See Results below)
 * 1) Transform again
 * 2) re-plate remaining 800μL of transformation from yesterday
 * 3) Start miniprep culture of single colony

Experimental

 * 1) Transformation of pTXB1+His QuikChange into DH10B
 * 2) 5μL QuikChange experimental reaction from  Friday (Daniel Catt's + 200μL  DH10B
 * 3) 5μL QuikChange control reaction from  Friday (Daniel Catt's + 200μL  DH10B
 * 4) * Follow standard transformation protocol, except increase heat-shock time to 45s
 * 5) * Plate 200μL on LBAmp and grow O/N at 37°C
 * 6) *&rarr; store remainder @ 4°C
 * 7) Re-plate transformation from  yesterday
 * 8) spread 800μL pTXB1+His/DH10B QuikChange experimental transformation from  yesterday on LBAmp plate
 * 9) spread 800μL pTXB1+His/DH10B QuikChange control transformation from  yesterday on LBAmp plate
 * 10) *&rarr; 37°C O/N
 * 11)  Miniprep
 * 12) 10 mL Muratore:Materials/Media/LB + 10 μL 100mg/mL  Amp + pTXB1+His/DH10B QuikChange experimental transformation colony from plate (colony #1)
 * 13) *&rarr; 37°C O/N with shaking

Results

 * Single colony on experimental plate from yesterday
 * &rarr; pTXB1His/DH10B QC colony #1
 * No colonies on control


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