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| | '''[[Montreal/Notebook/May|May 2008]]''' | '''[[Montreal/Notebook/June|June 2008]]''' | '''[[Montreal/Notebook/July|July 2008]]''' | '''[[Montreal/Notebook/August|August 2008]]''' | '''[[Montreal/Notebook/September|September 2008]]''' | '''[[Montreal/Notebook/October|October 2008]]''' | '''[[Montreal/Notebook/November|November 2008]]'''</center> | | '''[[Montreal/Notebook/May|May 2008]]''' | '''[[Montreal/Notebook/June|June 2008]]''' | '''[[Montreal/Notebook/July|July 2008]]''' | '''[[Montreal/Notebook/August|August 2008]]''' | '''[[Montreal/Notebook/September|September 2008]]''' | '''[[Montreal/Notebook/October|October 2008]]''' | '''[[Montreal/Notebook/November|November 2008]]'''</center> |
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| - | <p>'''June 2nd, 2008''':<ul>Growth of I-brick on culture</ul>
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| - | <ul>Midi-prep of both I and J brick followed by gel</ul>
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| - | <ul>Gel indicates no presence of DNA, will be confirmed by spectrophotometric assay</ul></p>
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| - | <p>'''June 3rd, 2008''':
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| - | <ul>Seeding of 5mL cultures of both I and J brick</ul>
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| - | <ul>Identity of colonies on I brick plates is suspect, must ensure that eventual DNA gel confirms exact restriction digest</ul></p>
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| - | <p>'''June 4th, 2008''':
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| - | <ul>Midiprep was done on the I and the J-brick. Once the isopropanol was added, the J-brick midiprep looked clear (no DNA was eluted). DNA gel needs to be done to confirm presence of DNA in both cases.</ul></p>
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| - | <p>'''June 9th, 2008''':
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| - | <ul>Seeded J and I-brick re-seeded for Maxi-Prep</ul>
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| - | <ul>Gel failed to confirm presence of previously collected DNA samples of J and I-brick, will be repeated following Maxi-Prep.</ul></p>
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| - | <p>'''June 10th, 2008''':
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| - | <ul>Since no growth was observed in the I-brick culture, the I brick was re-seeded.</ul>
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| - | <ul>The J-brick was diluted in 500-ml of LB broth(for a Maxiprep to be done the next day).</ul></p>
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| - | <p>'''June 11th, 2008''':<ul> Maxiprep of the J-brick.</ul>
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| - | <ul>Restriction digest of J-brick.</ul>
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| - | <ul>Dilution of I-brick in 500-ml of LB broth. </ul></p>
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| - | <p>'''June 12th, 2008''':<ul> Maxiprep of the I-brick.</ul>
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| - | <ul>Restriction digest and gel of June 11th and June 12th I-brick and J-brick DNA using EcoR1. Bands revealed at roughly 4000bp and 2500bp for I-brick (expected 2652bp and 3939bp). J brick single band that was not informative, new digestion to be completed tomorrow.</ul></p>
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| - |
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| - | <p>'''June 16th, 2008''':<ul> I-brick was seeded and diluted over last two days, but there was insufficient growth so it will be left to grow one more day before performing another midi-prep. This is to compliment the already successful Maxi-Prep that gave low concentrations of DNA.</ul>
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| - | <ul>Another gel was performed of previous J-brick preps that confirmed the absence of the desired plasmid, no DNA was detected when digested with EcoR1.</ul>
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| - | <ul>J-brick was re-transformed into TOP10 chemically competent cells and then plated on Amp+ plates.<ul/></p>
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