Team:Osaka/Notebook

From 2012.igem.org

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==Weekly Summaries==
==Weekly Summaries==
<h4>Week 1</h4>
<h4>Week 1</h4>
-
<p>Went through basic molecular biology lab skills and techniques with the new members. Extracted most of the existing parts required for our project from the iGEM 2012 Spring distribution plates, transformed and amplified the DNA for future use.</p>
+
<p>Went through basic molecular biology lab skills and techniques with the new members. Extracted most of the existing parts required for our project from the iGEM 2012 Spring distribution plates, transformed and amplified the DNA for future use.DNA extraction & purification (miniprep) of transformed parts, & gel runs to confirm lengths. Ligations to put together some common combinations (eg promoter + RBS).</p>
 +
<p>We transfered redioresistance parts from one vector(pSB1C3) to another vector(pSB1A3) and transformed plasmid DNA into Rosetta E.coli.
 +
</p>
<h4>Week 2</h4>
<h4>Week 2</h4>
-
<p>DNA extraction & purification (miniprep) of last week's transformed parts, & gel runs to confirm lengths. Sequencer was broken so we could not confirm sequence, but most parts' lengths turned out ok. Ligations to put together some common combinations (eg promoter + RBS).</p>
+
<p>We run the SDS-PAGE gel to check the expression of the gene products at the protein level. The result of DNA sequencing demonstrated that there is a mutation in PprA(Silent mutation).</p>
<h4>Week 3</h4>
<h4>Week 3</h4>
<p>Extraction & amplification of more basic parts from the distribution plates, plus ligation of a few more parts. Not much progress due to lab being closed for Obon holidays.</p>
<p>Extraction & amplification of more basic parts from the distribution plates, plus ligation of a few more parts. Not much progress due to lab being closed for Obon holidays.</p>
 +
<p>Completion of radioresistance parts and drafting of damage tolerance assay protocol.</p>
 +
 +
<h4>Week 4</h4>
 +
<p>We were going to assemble a promoter evaluation device utilizing firefly and renilla luciferase, but did not make  much progress due to low transformation efficiency(?).
 +
</p>
 +
 +
<h4>Week 5</h4>
 +
<p>Preliminary test (damage tolerance, Mitomycin C).</p>
 +
<p>We were going to assemble a promoter evaluation device utilizing firefly and renilla luciferase, but did not make much progress due to low transformation efficiency(?).
 +
</p>
 +
 +
<h4>Week 6</h4>
 +
<p>
 +
Damage tolerance assay (Mitomycin C): transformed cells were exposed to DNA damaging agents and then incubated on agar plates, and viability relative to control used as an evaluation of the abilities of PprI, PprA, PprM, RecA to confer resistance to radiation-induced DNA damage
 +
</p>
 +
 +
<h4>Week 7</h4>
 +
<p>
 +
Damage tolerance assay (Mitomycin C)</p>
 +
<p>
 +
Preliminary test (damage tolerance, Hydrogen peroxide)
 +
</p>
 +
 +
<h4>Week 8</h4>
 +
Damage tolerance assay (Hydrogen peroxide)
 +
 +
<h4>Week 9</h4>
 +
Damage tolerance assay (Hydrogen peroxide)
==Logbook Entries==
==Logbook Entries==
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|<span style={{{s12|"color:{{{c12|red}}};"}}}>12</span>
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|[[Team:Osaka/week3#August 12 (Sun)|12]]<span style={{{s12|"color:{{{c12|red}}};"}}}></span>
|<span style={{{s13|"color:{{{c13|red}}};"}}}>13</span>
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|<span style={{{s14|"color:{{{c14|red}}};"}}}>14</span>
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|<span style={{{s15|"color:{{{c15|red}}};"}}}>15</span>
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|colspan="6"|
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Latest revision as of 13:45, 25 September 2012

Weekly Summaries

Week 1

Went through basic molecular biology lab skills and techniques with the new members. Extracted most of the existing parts required for our project from the iGEM 2012 Spring distribution plates, transformed and amplified the DNA for future use.DNA extraction & purification (miniprep) of transformed parts, & gel runs to confirm lengths. Ligations to put together some common combinations (eg promoter + RBS).

We transfered redioresistance parts from one vector(pSB1C3) to another vector(pSB1A3) and transformed plasmid DNA into Rosetta E.coli.

Week 2

We run the SDS-PAGE gel to check the expression of the gene products at the protein level. The result of DNA sequencing demonstrated that there is a mutation in PprA(Silent mutation).

Week 3

Extraction & amplification of more basic parts from the distribution plates, plus ligation of a few more parts. Not much progress due to lab being closed for Obon holidays.

Completion of radioresistance parts and drafting of damage tolerance assay protocol.

Week 4

We were going to assemble a promoter evaluation device utilizing firefly and renilla luciferase, but did not make much progress due to low transformation efficiency(?).

Week 5

Preliminary test (damage tolerance, Mitomycin C).

We were going to assemble a promoter evaluation device utilizing firefly and renilla luciferase, but did not make much progress due to low transformation efficiency(?).

Week 6

Damage tolerance assay (Mitomycin C): transformed cells were exposed to DNA damaging agents and then incubated on agar plates, and viability relative to control used as an evaluation of the abilities of PprI, PprA, PprM, RecA to confer resistance to radiation-induced DNA damage

Week 7

Damage tolerance assay (Mitomycin C)

Preliminary test (damage tolerance, Hydrogen peroxide)

Week 8

Damage tolerance assay (Hydrogen peroxide)

Week 9

Damage tolerance assay (Hydrogen peroxide)

Logbook Entries

Click on any day in the calendar below to display detailed logbook entry.

August
Week S M T W T F S
week1 1 2 3 4
week2 5 6 7 8 9 10 11
week3 12 13 14 15 16 17 18
week4 19 20 21 22 23 24 25
week5 26 27 28 29 30 31
September
Week S M T W T F S
week5 1
week6 2 3 4 5 6 7 8
week7 9 10 11 12 13 14 15
week8 16 17 18 19 20 21 22
week9 23 24 25 26 27 28 29
Retrieved from "http://2012.igem.org/Team:Osaka/Notebook"