Team:Montreal/Notebook

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==Lab Progress==
==Lab Progress==
-
'''May 21st, 2008''': <ul>Resctriction enzyme digest was done on the J-brick with EcoRI. Gel was run on the J-brick after the restriction digest. No DNA was detected on the gel (the ladder was visible on the gel).</ul>
+
<p>'''May 21st, 2008''': <ul>Resctriction enzyme digest was done on the J-brick with EcoRI. Gel was run on the J-brick after the restriction digest. No DNA was detected on the gel (the ladder was visible on the gel).</ul></p>
-
 
+
<p>'''May 22nd, 2008''': <ul>Prepared TOP10 Competent cells for eventual transformation.</ul>
-
'''May 22nd, 2008''': <ul>Prepared TOP10 Competent cells for eventual transformation.</ul>
+
<ul>Performed Mini-prep on Reporter+ Cells</ul>
<ul>Performed Mini-prep on Reporter+ Cells</ul>
-
<ul>Performed Digest and Gel on Reporter plasmid extract -- no DNA present, suggest follow-up maxi-prep</ul>
+
<ul>Performed Digest and Gel on Reporter plasmid extract -- no DNA present, suggest follow-up maxi-prep</ul></p>
 +
<p>'''May 23rd, 2008''': <ul>Transformed Top10 cells with Puc19 to ensure that the competent cell procedure was successful. Growth was observed, therefore procedure was successful.</ul></p>
-
'''May 23rd, 2008''': <ul>Transformed Top10 cells with Puc19 to ensure that the competent cell procedure was successful. Growth was observed, therefore procedure was successful.</ul>
+
<p>'''May 25th, 2008''': <ul>Diluted Reporter cells 1/1000 with 5ul Kan/mL culture for 16h incubation at 5:30pm. To be used for Maxiprep at 9:30am-1:30pm.</ul></p>
 +
<p>'''May 28th, 2008''':<ul> Ran gel on Elowitz reporter DNA cut with EcoR1; 2 bands</ul>
 +
<ul>0.7 kb and 2.0 kb, confirms identity of reporter DNA.</ul>
 +
<ul>Seeded syn-I and J-40001 into amp/kan LB and kan LB.</ul></p>
-
'''May 25th, 2008''': <ul>Diluted Reporter cells 1/1000 with 5ul Kan/mL culture for 16h incubation at 5:30pm. To be used for Maxiprep at 9:30am-1:30pm.</ul>
 
-
 
-
 
-
'''May 28th, 2008''':<ul> Ran gel on Elowitz reporter DNA cut with EcoR1; 2 bands</ul>
 
-
<ul>0.7 kb and 2.0 kb, confirms identity of reporter DNA.</ul>
 
-
<ul>Seeded syn-I and J-40001 into amp/kan LB and kan LB.</ul>
 
<p>'''May 29th, 2008''': <ul>Growth of J-brick in culture - No growth of I-brick on culture</ul>
<p>'''May 29th, 2008''': <ul>Growth of J-brick in culture - No growth of I-brick on culture</ul>
<ul> Seeded J-brick for Midi-Prep in 40mL LB with ampicillin</ul>
<ul> Seeded J-brick for Midi-Prep in 40mL LB with ampicillin</ul>
-
<ul>Transformed TOP10 cells with both I brick and Reporter Plasmid</ul>
+
<ul>Transformed TOP10 cells with both I brick and Reporter Plasmid</ul></p>
-
 
+
<p>'''June 2nd, 2008''':<ul>Growth of I-brick on culture</ul>
-
'''June 2nd, 2008''':<ul>Growth of I-brick on culture</ul>
+
<ul>Midi-prep of both I and J brick followed by gel</ul>
<ul>Midi-prep of both I and J brick followed by gel</ul>
-
<ul>Gel indicates no presence of DNA, will be confirmed by spectrophotometric assay</ul>
+
<ul>Gel indicates no presence of DNA, will be confirmed by spectrophotometric assay</ul></p>
-
 
+
<p>'''June 3rd, 2008''':
-
'''June 3rd, 2008''':
+
<ul>Seeding of 5mL cultures of both I and J brick</ul>
<ul>Seeding of 5mL cultures of both I and J brick</ul>
-
<ul>Identity of colonies on I brick plates is suspect, must ensure that eventual DNA gel confirms exact restriction digest</ul>
+
<ul>Identity of colonies on I brick plates is suspect, must ensure that eventual DNA gel confirms exact restriction digest</ul></p>
 +
<p>'''June 4th, 2008''':
 +
<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>
-
'''June 4th, 2008''':
+
<p>'''June 9th, 2008''':
-
<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>
+
-
 
+
-
 
+
-
'''June 9th, 2008''':
+
<ul>Seeded J and I-brick re-seeded for Maxi-Prep</ul>
<ul>Seeded J and I-brick re-seeded for Maxi-Prep</ul>
-
<ul>Gel failed to confirm presence of previously collected DNA samples of J and I-brick, will be repeated following Maxi-Prep.</ul>
+
<ul>Gel failed to confirm presence of previously collected DNA samples of J and I-brick, will be repeated following Maxi-Prep.</ul></p>
-
 
+
<p>'''June 10th, 2008''':
-
'''June 10th, 2008''':
+
<ul>Since no growth was observed in the I-brick culture, the I brick was re-seeded.</ul>
<ul>Since no growth was observed in the I-brick culture, the I brick was re-seeded.</ul>
-
<ul>The J-brick was diluted in 500-ml of LB broth(for a Maxiprep to be done the next day).</ul>
+
<ul>The J-brick was diluted in 500-ml of LB broth(for a Maxiprep to be done the next day).</ul></p>
-
 
+
-
'''June 11th, 2008''':<ul> Maxiprep of the J-brick.</ul>
+
<p>'''June 11th, 2008''':<ul> Maxiprep of the J-brick.</ul>
<ul>Restriction digest of J-brick.</ul>
<ul>Restriction digest of J-brick.</ul>
-
<ul>Dilution of I-brick in 500-ml of LB broth. </ul>
+
<ul>Dilution of I-brick in 500-ml of LB broth. </ul></p>

Revision as of 02:49, 12 June 2008

Home The Team The Project Parts Submitted to the Registry Modeling Notebook

Lab Progress

May 21st, 2008:

    Resctriction enzyme digest was done on the J-brick with EcoRI. Gel was run on the J-brick after the restriction digest. No DNA was detected on the gel (the ladder was visible on the gel).

May 22nd, 2008:

    Prepared TOP10 Competent cells for eventual transformation.
    Performed Mini-prep on Reporter+ Cells
    Performed Digest and Gel on Reporter plasmid extract -- no DNA present, suggest follow-up maxi-prep

May 23rd, 2008:

    Transformed Top10 cells with Puc19 to ensure that the competent cell procedure was successful. Growth was observed, therefore procedure was successful.

May 25th, 2008:

    Diluted Reporter cells 1/1000 with 5ul Kan/mL culture for 16h incubation at 5:30pm. To be used for Maxiprep at 9:30am-1:30pm.

May 28th, 2008:

    Ran gel on Elowitz reporter DNA cut with EcoR1; 2 bands
    0.7 kb and 2.0 kb, confirms identity of reporter DNA.
    Seeded syn-I and J-40001 into amp/kan LB and kan LB.

May 29th, 2008:

    Growth of J-brick in culture - No growth of I-brick on culture
    Seeded J-brick for Midi-Prep in 40mL LB with ampicillin
    Transformed TOP10 cells with both I brick and Reporter Plasmid

June 2nd, 2008:

    Growth of I-brick on culture
    Midi-prep of both I and J brick followed by gel
    Gel indicates no presence of DNA, will be confirmed by spectrophotometric assay

June 3rd, 2008:

    Seeding of 5mL cultures of both I and J brick
    Identity of colonies on I brick plates is suspect, must ensure that eventual DNA gel confirms exact restriction digest

June 4th, 2008:

    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.

June 9th, 2008:

    Seeded J and I-brick re-seeded for Maxi-Prep
    Gel failed to confirm presence of previously collected DNA samples of J and I-brick, will be repeated following Maxi-Prep.

June 10th, 2008:

    Since no growth was observed in the I-brick culture, the I brick was re-seeded.
    The J-brick was diluted in 500-ml of LB broth(for a Maxiprep to be done the next day).

June 11th, 2008:

    Maxiprep of the J-brick.
    Restriction digest of J-brick.
    Dilution of I-brick in 500-ml of LB broth.

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