Team:Montreal/Notebook

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Revision as of 02:39, 12 June 2008

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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:

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.

@@ Line 10: / Line 10: @@
 ==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 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>
-'''May 22nd, 2008''': - Prepared TOP10 Competent cells for eventual transformation.
+'''May 22nd, 2008''': <ul>Prepared TOP10 Competent cells for eventual transformation.</ul>
-- Performed Mini-prep on Reporter+ Cells
+<ul>Performed Mini-prep on Reporter+ Cells</ul>
-- Performed Digest and Gel on Reporter plasmid extract -- no DNA present, suggest follow-up maxi-prep
+<ul>Performed Digest and Gel on Reporter plasmid extract -- no DNA present, suggest follow-up maxi-prep</ul>
-'''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 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>
-'''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 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''': - Ran gel on Elowitz reporter DNA cut with EcoR1; 2 bands - 0.7 kb and 2.0 kb, confirms identity of reporter DNA.
+'''May 28th, 2008''':<ul> Ran gel on Elowitz reporter DNA cut with EcoR1; 2 bands</ul>
-- Seeded syn-I and J-40001 into amp/kan LB and kan LB.
+<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>
-'''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
+'''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>Transformed TOP10 cells with both I brick and Reporter Plasmid</ul>
-'''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 2nd, 2008''':<ul>Growth of I-brick on culture</ul>
+<ul>Midi-prep of both I and J brick followed by gel</ul>
-'''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
+<ul>Gel indicates no presence of DNA, will be confirmed by spectrophotometric assay</ul>
+<p>'''June 3rd, 2008''':
+<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>
 '''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.