User:Kathryn Muratore/Notebook/AU CHEM-570 lab prep/2011/06/16

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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) Glycerol stocks
 * 2) * 750μL pTXB1His/DH10B QC colony #2 culture from yesterday + 250μL 60% sterile glycerol &rarr;  tube #70
 * 3) * 750μL pTXB1His/DH10B QC colony #3 culture from yesterday + 250μL 60% sterile glycerol  &rarr;  tube #71
 * 4) * 750μL pTXB1/DH10B colony #1 culture from yesterday + 250μL 60% sterile glycerol  &rarr;  tube #72
 * 5) *&rarr; -80°C
 * 6)  Miniprep O/N cultures from  yesterday
 * 7) * Cultures grew about 17h
 * 8) * Followed standard miniprep protocol
 * 9) Quantify [minipreps]
 * 10) * Done with Daniel Catt
 * 11) 95μL water + 5μL DNA from miniprep
 * 12) *&rarr; read A260 and A280
 * 13) NheI digest
 * 14) * performing sequential double digest, so use optimal buffer for SapI instead of NEB buffer 2
 * 15) * modify NheI digest protocol as follows
 * &rarr; 2h @ 37°C
 * &rarr; 20' @ 65°C (it took awhile to get the block to the right temp, so they were on ice for a bit, then at ~70°C before the block reached 65°C)
 * Note: I should have done a PCR cleanup on the BSA PCR sample! I will repeat this. *Kathryn Muratore 10:20, 20 June 2011 (EDT):
 * In retrospect, I'm not sure that I even used the correct tube here *Kathryn Muratore 16:26, 20 June 2011 (EDT):
 * I did use the right tube, as seen on the gels later that week *Kathryn Muratore 10:33, 28 June 2011 (EDT):
 * &rarr; store @ -20°C

Results

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