Team:Montreal/Notebook

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Latest revision as of 17:22, 1 August 2008

Home	The Team	The Project	Parts	Notebook	Modeling	Links	Sponsors

Lab Protocols

Key Elements of the Central Dogma of cloning

1. Restriction Digest

2. Making a DNA Agarose Gel

3. Gel extraction

4. DNA quantity assessment

5. Ligation

6. Cell Transformation

7. Seeding method

8. DNA Extraction: Mini-, Midi-, Maxiprep

Lab Progress

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-{| style="color:#ffffff;background-color:#cd0000;" cellpadding="3" cellspacing="5" border="2" bordercolor="#ffffff" width="75%" align="center"
+{|style="font color="#ffffff"; background-color:#cd0000; cellpadding="3" cellspacing="5" border="2" bordercolor="#cd0000"border-spacing:6px; text-align:center" width="960px"
-!align="center"|[[Team:Montreal|Home]]
+!style="text-align:center; background-color:#cd0000; border-width:0px; padding:3px;"|[[Team:Montreal|<font color="#ffffff">Home</font>]]
-!align="center"|[[Team:Montreal/Team|The Team]]
+!style="text-align:center; background-color:#cd0000; border-width:0px; padding:3px;"|[[Team:Montreal/Team|<font color="#ffffff">The Team</font>]]
-!align="center"|[[Team:Montreal/Project|The Project]]
+!style="text-align:center; background-color:#cd0000; border-width:0px; padding:3px;"|[[Team:Montreal/Project|<font color="#ffffff">The Project</font>]]
-!align="center"|[[Team:Montreal/Parts|Parts Submitted to the Registry]]
+!style="text-align:center; background-color:#cd0000; border-width:0px; padding:3px;"|[[Team:Montreal/Parts|<font color="#ffffff">Parts</font>]]
-!align="center"|[[Team:Montreal/Modeling|Modeling]]
+!style="text-align:center; background-color:#cd0000; border-width:0px; padding:3px;"|[[Team:Montreal/Notebook|<font color="#ffffff">Notebook</font>]]
-!align="center"|[[Team:Montreal/Notebook|Notebook]]
+!style="text-align:center; background-color:#cd0000; border-width:0px; padding:3px;"|[[Team:Montreal/Modeling|<font color="#ffffff">Modeling</font>]]
+!style="text-align:center; background-color:#cd0000; border-width:0px; padding:3px;"|[[Team:Montreal/Links|<font color="#ffffff">Links</font>]]
+!style="text-align:center; background-color:#cd0000; border-width:0px; padding:3px;"|[[Team:Montreal/Sponsors|<font color="#ffffff">Sponsors</font>]]
 |}
-==Lab Progress==
-<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>
+==Lab Protocols==
-<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></p>
+{| align="right"
-<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>
+|align="right" |[[Image:Central_dogma.jpg]]
-<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>
+Key Elements of the Central Dogma of cloning
-<ul>Seeded syn-I and J-40001 into amp/kan LB and kan LB.</ul></p>
-<p>'''May 29th, 2008''': <ul>Growth of J-brick in culture - No growth of I-brick on culture</ul>
+. [https://2008.igem.org/Team:Montreal/Restriction_digest Restriction Digest]
-<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></p>
+. [https://2008.igem.org/Team:Montreal/DNA_Agarose_Gel Making a DNA Agarose Gel]
-<p>'''June 2nd, 2008''':<ul>Growth of I-brick on culture</ul>
-<ul>Midi-prep of both I and J brick followed by gel</ul>
+. [https://2008.igem.org/Team:Montreal/Gel_extraction Gel extraction]
-<ul>Gel indicates no presence of DNA, will be confirmed by spectrophotometric assay</ul></p>
-<p>'''June 3rd, 2008''':
+. [https://2008.igem.org/Team:Montreal/Confirmation_quantity DNA quantity assessment]
-<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>
+. [https://2008.igem.org/Team:Montreal/Ligation Ligation]
-<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>
+. [https://2008.igem.org/Team:Montreal/Cell_transformation Cell Transformation]
-<p>'''June 9th, 2008''':
-<ul>Seeded J and I-brick re-seeded for Maxi-Prep</ul>
+. [https://2008.igem.org/Team:Montreal/Seeding Seeding method]
-<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''':
+. [https://2008.igem.org/Team:Montreal/DNA_Extraction DNA Extraction: Mini-, Midi-, Maxiprep]
-<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></p>
+==Lab Progress==
-<p>'''June 11th, 2008''':<ul> Maxiprep of the J-brick.</ul>
+<p><center>
-<ul>Restriction digest of J-brick.</ul>
+'''[[Team:Montreal/Notebook/May|May 2008]]''' | '''[[Team:Montreal/Notebook/June|June 2008]]''' | '''[[Team:Montreal/Notebook/July|July 2008]]''' | '''[[Team:Montreal/Notebook/August|August 2008]]''' | '''[[Team:Montreal/Notebook/September|September 2008]]''' | '''[[Team:Montreal/Notebook/October|October 2008]]''' | '''[[Team:Montreal/Notebook/November|November 2008]]'''</center><br>
-<ul>Dilution of I-brick in 500-ml of LB broth. </ul>