Team:CBNU-Korea/Notebook

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{{:Template:CBNU-Korea}}
{{:Template:CBNU-Korea}}
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<div id="box" style="width: 700px; margin-left: 137px; padding: 5px; border: 3px solid #000; background-color: #fe2b33;">
 
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<div id="template" style="text-align: center; font-weight: bold; font-size: large; color: #f6f6f6; padding: 5px;">
 
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This is a template page. READ THESE INSTRUCTIONS.
 
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</div>
 
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<div id="instructions" style="text-align: center; font-weight: normal; font-size: small; color: #f6f6f6; padding: 5px;">
 
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You are provided with this team page template with which to start the iGEM season.  You may choose to personalize it to fit your team but keep the same "look." Or you may choose to take your team wiki to a different level and design your own wiki.  You can find some examples <a href="https://2008.igem.org/Help:Template/Examples">HERE</a>.
 
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</div>
 
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<div id="warning" style="text-align: center; font-weight: bold; font-size: small; color: #f6f6f6; padding: 5px;">
 
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You <strong>MUST</strong> have a team description page, a project abstract, a complete project description, a lab notebook, and a safety page.  PLEASE keep all of your pages within your teams namespace. 
 
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</div>
 
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</div>
 
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</html>
 
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<!-- *** End of the alert box *** -->
 
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+
<p>5/7<br>
-
{|align="justify"
+
  <strong>Membership meeting</strong><br>
-
|You can write a background of your team here.  Give us a background of your team, the members, etc.  Or tell us more about something of your choosing.
+
  New participant of CBNU-KOREA team.<br>
-
|[[Image:CBNU-Korea_logo.png|200px|right|frame]]
+
  Introduced what is the iGEM, what we should do if we join the competition.<br>
-
|-
+
  5/15<br>
-
|
+
  <strong>Concept meeting</strong><br>
-
''Tell us more about your project.  Give us background.  Use this is the abstract of your project.  Be descriptive but concise (1-2 paragraphs)''
+
  Decided 2010 CBNU-KOREA iGEM team’s project(almost 4hr)<br>
-
|[[Image:CBNU-Korea_team.png|right|frame|Your team picture]]
+
  5/18<br>
-
|-
+
  <strong>Researching </strong><br>
-
|
+
  Finding related articles, paper and so on.<br>
-
|align="center"|[[Team:CBNU-Korea | Team Example]]
+
  Essential gene data search<br>
-
|}
+
  5/25<br>
-
 
+
  <strong>Strategy about database</strong><br>
-
<!--- The Mission, Experiments --->
+
  Planned how to re-construct essential gene database.<br>
-
 
+
  5/27<br>
-
{| style="color:#1b2c8a;background-color:#0c6;" cellpadding="3" cellspacing="1" border="1" bordercolor="#fff" width="62%" align="center"
+
  <strong>Gathering</strong><br>
-
!align="center"|[[Team:CBNU-Korea|Home]]
+
  Gathering  the essential genes data from DEG, NCBI.<br>
-
!align="center"|[[Team:CBNU-Korea/Team|Team]]
+
  Starting  to lean about experimental technique.<br>
-
!align="center"|[https://igem.org/Team.cgi?year=2010&team_name=CBNU-Korea Official Team Profile]
+
  6/1<br>
-
!align="center"|[[Team:CBNU-Korea/Project|Project]]
+
  Clean  up the lab room. Too dirty…<br>
-
!align="center"|[[Team:CBNU-Korea/Parts|Parts Submitted to the Registry]]
+
  Computer  engineering department meeting about project<br>
-
!align="center"|[[Team:CBNU-Korea/Modeling|Modeling]]
+
  6/24<br>
-
!align="center"|[[Team:CBNU-Korea/Notebook|Notebook]]
+
  Summer  iGEM Workshop Asia <br>
-
!align="center"|[[Team:CBNU-Korea/Safety|Safety]]
+
  7/19<br>
-
|}
+
  <strong>Plasmid Culture</strong> <br>
-
 
+
  · Pick up a single colony from fresh cultured LB agar plate.<br>
-
 
+
  · Our established media and inoculate the cell into the 5ml of fresh LB liquid media (contains antibiotics) and our established media at 37℃ with shaking for 12hr.<br>
-
5/7
+
  · plasmid : pSB1C3, pSB1A3, pSB1T3, pSB1K3 (contain J54450)<br>
-
Membership meeting
+
  7/20<br>
-
New participant of CBNU-KOREA team.
+
  <strong>Plasmid extract</strong> <br>
-
Introduced what is the iGEM, what we should do if we join the competition.
+
  · Uses kit and extracted plasmid. (pSB1C3, pSB1A3, pSB1T3, pSB1K3)<br>
-
5/15
+
  · Confirmed size by electrophoresis.<br>
-
Concept meeting
+
  7/23<br>
-
Decided 2010 CBNU-KOREA iGEM team’s project(almost 4hr)
+
  <strong>Plasmid Digest</strong> <br>
-
5/18
+
  · Digest of 4 plasmid.<br>
-
Researching  
+
  · Used EcoRⅠ-PstⅠ restriction endonuclease.<br>
-
Finding related articles, paper and so on.
+
  · Confirmed size by electrophoresis.<br>
-
Essential gene data search
+
  7/24<br>
-
5/25
+
  <strong>Primer Design </strong> <br>
-
Strategy about database
+
  · Designed <em>Vibrio cholerae</em>'s <em>oriC2vc, parA, parB, parS, dif</em> gene primer using ‘Ginko primer’.<br>
-
Planned how to re-construct essential gene database.
+
  7/28<br>
-
7/19
+
  <strong>DNA Amplification</strong> <br>
-
Plasmid Culture
+
  · Amplified <em>ori, parA, parB, parS, dif</em> gene of <em>vibrio cholerae</em>.<br>
-
• Pick up a single colony from fresh cultured LB agar plate.
+
  · Confirmed size by electrophoresis. And that’s correct.<br>
-
• Our established media and inoculate the cell into the 5ml of fresh LB liquid media (contains antibiotics) and our established media at 37℃ with shaking for 12hr.
+
  <strong>Concentration</strong> <br>
-
• plasmid : pSB1C3, pSB1A3, pSB1T3, pSB1K3 (contain J54450)
+
  · Because concentration of <em>parS, dif</em> gene was low, it was hard to confirm result by electrophoresis.<br>
-
7/20
+
  · Concentrated gene sample<br>
-
Plasmid extract
+
  7/29<br>
-
• Uses kit and extracted plasmid. (pSB1C3, pSB1A3, pSB1T3, pSB1K3)
+
  <strong>DNA(gene) Digest</strong> <br>
-
• Confirmed size by electrophoresis.
+
  · Digest of 4 plasmid.<br>
-
7/23
+
  · Used EcoRⅠ-PstⅠ restriction endonuclease.<br>
-
Plasmid Digest
+
  · Confirmed size by electrophoresis.<br>
-
• Digest of 4 plasmid.
+
  7/30<br>
-
• Used EcoRⅠ-PstⅠ restriction endonuclease.
+
  <strong>Prepared Compentent cell</strong> <br>
-
• Confirmed size by electrophoresis.
+
  · <em>E.coli</em> DH10B, DH5α, JM109<br>
-
7/24
+
  8/2<br>
-
Primer Design  
+
  <strong>DNA Ligation/Transformation</strong> <br>
-
• Designed Vibrio cholerae's oriC2vc, parA, parB, parS, dif gene primer using ‘Ginko primer’.
+
  · Ligation inserting gene in vector<br>
-
7/28
+
  (<em>parA</em>+pSB1A3, <em>parS</em>+pSB1C3, <em>ori</em>+pSB1K3, <em>dif</em>+pSB1A3)<br>
-
DNA Amplification
+
  · Transformation (Competent cell : <em>E.coli </em>DH10B)<br>
-
• Amplified ori, parA, parB, parS, dif gene of vibrio cholerae.
+
  · Spreading on plate (each antibiotics contained LB agar media)<br>
-
• Confirmed size by electrophoresis.
+
  8/3<br>
-
Concentration
+
  <strong>Cell Culture</strong> <br>
-
• Because concentration of parS, dif gene was low, it was hard to confirm result by electrophoresis.
+
  · Pick up a single colony from fresh cultured LB agar plate.<br>
-
• Concentrated gene sample
+
  · Our established media and inoculate the cell into the 5ml of fresh LB liquid media (contains antibiotics) and our established media at 37℃ with shaking for 12hr.<br>
-
7/29
+
  · vector : <em>parA</em>+pSB1A3, <em>parS</em>+pSB1C3, <em>ori</em>+pSB1K3, <em>dif</em>+pSB1A3<br>
-
DNA(gene) Digest
+
  8/4<br>
-
• Digest of 4 plasmid.
+
  <strong>Vector extract</strong> <br>
-
• Used EcoRⅠ-PstⅠ restriction endonuclease.
+
  · Uses kit and extracted plasmid.<br>
-
• Confirmed size by electrophoresis.
+
  (parA+pSB1A3, parS+pSB1C3, ori+pSB1K3, dif+pSB1A3)<br>
-
7/30
+
  · Confirmed size by electrophoresis.<br>
-
Prepared Compentent cell
+
  8/9<br>
-
• E.coli DH10B, DH5α, JM109
+
  <strong>Plasmid Digest</strong> <br>
-
8/2
+
  · Digest of 4 vector.<br>
-
DNA Ligation/Transformation
+
  · ori+pSB1K3, parA+pSB1A3, parS+pSB1C3 digests by EcoRⅠ-SpeⅠ restriction endonuclease and dif+pSB1A3 gene digested by XbaⅠ-PstⅠ restriction endonuclease.<br>
-
• Ligation inserting gene in vector
+
  · Confirmed size by electrophoresis.<br>
-
(parA+pSB1A3, parS+pSB1C3, ori+pSB1K3, dif+pSB1A3)
+
  · Came out as size of <em>ori</em>+pSB1K3 gene is smaller.<br>
-
• Transformation (Competent cell : E.coli DH10B)
+
  8/10<br>
-
• Spreading on plate (each antibiotics contained LB agar media)
+
  <strong>Sequencing Order</strong> <br>
-
8/3
+
  · Analyzed plasmid DNA extraction (cultured cell)<br>
-
Cell Culture
+
  8/16<br>
-
• Pick up a single colony from fresh cultured LB agar plate.
+
  <strong>Primer Again design </strong> <br>
-
• Our established media and inoculate the cell into the 5ml of fresh LB liquid media (contains antibiotics) and our established media at 37℃ with shaking for 12hr.
+
  · According to analysis result, primer of <em>ori</em> gene knew wrong fact.<br>
-
• vector : parA+pSB1A3, parS+pSB1C3, ori+pSB1K3, dif+pSB1A3
+
  · Designed primer of <em>ori</em> gene again.<br>
-
8/4
+
  · Primer of I51020 and p1003 designed.<br>
-
Vector extract
+
  8/20<br>
-
• Uses kit and extracted plasmid.
+
  <strong>DNA Amplification</strong> <br>
-
(parA+pSB1A3, parS+pSB1C3, ori+pSB1K3, dif+pSB1A3)
+
  · Amplified I51020, p1003<br>
-
• Confirmed size by electrophoresis.
+
  · Confirmed size by electrophoresis.<br>
-
8/9
+
  8/23<br>
-
Plasmid Digest
+
  <strong>Plasmid Digest</strong> <br>
-
• Digest of 4 vector.
+
  · I51020 digests by EcoRⅠ-PstⅠ restriction endonuclease.<br>
-
• ori+pSB1K3, parA+pSB1A3, parS+pSB1C3 digests by EcoRⅠ-SpeⅠ restriction endonuclease and dif+pSB1A3 gene digested by XbaⅠ-PstⅠ restriction endonuclease.
+
  · p1003 digests by XbaⅠ-PstⅠ restriction endonuclease.<br>
-
• Confirmed size by electrophoresis.
+
  · Confirmed size by electrophoresis.<br>
-
• Came out as size of ori+pSB1K3 gene is smaller.
+
  8/24<br>
-
8/10
+
  <strong>DNA Ligation/Transformation</strong> <br>
-
Sequencing Order
+
  · Ligated inserting gene in vector<br>
-
• Analyzed plasmid DNA extraction (cultured cell)
+
  (Inserted <em>ori</em> gene and p1003 antibiotic resistance cassette in I51020 vector, inserted <em>parS </em>gene and <em>dif </em>gene in pSB1A3 vector)<br>
-
8/16
+
  · Transformation (Competent cell : <em>E.coli </em>DH10B)<br>
-
Primer Again design  
+
  · Spreading on plate (each antibiotics contained LB agar media)</p>
-
• According to analysis result, primer of ori gene knew wrong fact.
+
<p>8/25<br>
-
• Designed primer of ori gene again.
+
  I51020 transformation failed<br>
-
• Primer of I51020 and p1003 designed.
+
  Plan to design Cn vector <br>
-
8/20
+
  Ready to P1003, <em>ori</em> miniprep </p>
-
DNA Amplification
+
<p>9/2</p>
-
• Amplified I51020, p1003
+
<p>Primer design for Cn vector. May be it won’t  work….</p>
-
• Confirmed size by electrophoresis.
+
<p>All parts re-transformation for selection  and miniprep.</p>
-
8/23
+
<p><strong>&nbsp;</strong></p>
-
Plasmid Digest
+
<p>9/7 </p>
-
• I51020 digests by EcoRⅠ-PstⅠ restriction endonuclease.
+
<p>Cn vector primer arrived.<br>
-
• p1003 digests by XbaⅠ-PstⅠ restriction endonuclease.
+
  Ready to PSB1C3 plasmid miniprep<br>
-
• Confirmed size by electrophoresis.
+
  <em>Ori</em> X/S digestion<strong>. </strong></p>
-
8/24
+
<p><strong>&nbsp;</strong></p>
-
DNA Ligation/Transformation
+
<p><strong>9/10</strong></p>
-
• Ligated inserting gene in vector
+
<p><strong>Cn  vector PCR, purification, ligation.</strong><br>
-
(Inserted ori gene and p1003 antibiotic resistance cassette in I51020 vector, inserted parS gene and dif gene in pSB1A3 vector)
+
  <strong>Cn  vector PCR Product electroporhesis.</strong><br>
-
• Transformation (Competent cell : E.coli DH10B)
+
  <strong>9/15</strong><br>
-
• Spreading on plate (each antibiotics contained LB agar media)
+
  <strong>Cn  vector transformation failed.</strong><br>
 +
  <strong>Cn  vercor PCR condition re arrangement</strong><br>
 +
  <strong>PrctB  promoter primer design</strong></p>
 +
<p><strong>&nbsp;</strong></p>
 +
<p><strong>9/25</strong><br>
 +
  <strong>Cn  vector + <em>or</em>i(X-S) ligation,  transformation.</strong><br>
 +
  <strong>PrctB  primer arrived , PCR </strong></p>
 +
<p><strong>9/27</strong><br>
 +
  <strong>Cn  vector colony appear. But not red, may be failure.</strong><br>
 +
  <strong>PrctB  + rctB ligation.</strong><br>
 +
  <strong>Cn  vector transformation</strong></p>
 +
<p><strong>9/30 </strong><br>
 +
  <strong>Cn  vector colony appear again. Not red too. Ready to miniprep.</strong><br>
 +
  <strong>parB  + GFP fusion </strong></p>
 +
<p><strong>10/3</strong><br>
 +
  <strong>parA,  parS, dif ligation PSB1C3 and PSB1A3</strong><br>
 +
  <strong>computer  engineering student meeting,</strong></p>
 +
<p><strong>&nbsp;</strong></p>
 +
<p><strong>10/8</strong><br>
 +
  <strong>Ori  PCR and miniprep</strong><br>
 +
  <strong>Ori  digestion EP/XP/XS</strong></p>
 +
<p><strong>&nbsp;</strong></p>
 +
<p><strong>10/10</strong></p>
 +
<p><strong>parS,  dif parts complete.</strong><br>
 +
  <strong>parA  re PCR.</strong></p>
 +
<p><strong>10/15</strong><br>
 +
  <strong>All  members of CBNU-KOREA team meeting</strong></p>
 +
<p><strong>10/21</strong><br>
 +
  <strong>Parts  making start.</strong></p>
 +
<p><strong>Ori  function is not checked.</strong></p>
 +
<p><strong>10/25</strong></p>
 +
<p><strong>Parts  sending </strong><br>
 +
  <strong>Prepare  Team wiki freeze</strong></p>

Latest revision as of 03:22, 28 October 2010


5/7
Membership meeting
New participant of CBNU-KOREA team.
Introduced what is the iGEM, what we should do if we join the competition.
5/15
Concept meeting
Decided 2010 CBNU-KOREA iGEM team’s project(almost 4hr)
5/18
Researching
Finding related articles, paper and so on.
Essential gene data search
5/25
Strategy about database
Planned how to re-construct essential gene database.
5/27
Gathering
Gathering the essential genes data from DEG, NCBI.
Starting to lean about experimental technique.
6/1
Clean up the lab room. Too dirty…
Computer engineering department meeting about project
6/24
Summer iGEM Workshop Asia
7/19
Plasmid Culture
· Pick up a single colony from fresh cultured LB agar plate.
· Our established media and inoculate the cell into the 5ml of fresh LB liquid media (contains antibiotics) and our established media at 37℃ with shaking for 12hr.
· plasmid : pSB1C3, pSB1A3, pSB1T3, pSB1K3 (contain J54450)
7/20
Plasmid extract
· Uses kit and extracted plasmid. (pSB1C3, pSB1A3, pSB1T3, pSB1K3)
· Confirmed size by electrophoresis.
7/23
Plasmid Digest
· Digest of 4 plasmid.
· Used EcoRⅠ-PstⅠ restriction endonuclease.
· Confirmed size by electrophoresis.
7/24
Primer Design
· Designed Vibrio cholerae's oriC2vc, parA, parB, parS, dif gene primer using ‘Ginko primer’.
7/28
DNA Amplification
· Amplified ori, parA, parB, parS, dif gene of vibrio cholerae.
· Confirmed size by electrophoresis. And that’s correct.
Concentration
· Because concentration of parS, dif gene was low, it was hard to confirm result by electrophoresis.
· Concentrated gene sample
7/29
DNA(gene) Digest
· Digest of 4 plasmid.
· Used EcoRⅠ-PstⅠ restriction endonuclease.
· Confirmed size by electrophoresis.
7/30
Prepared Compentent cell
· E.coli DH10B, DH5α, JM109
8/2
DNA Ligation/Transformation
· Ligation inserting gene in vector
(parA+pSB1A3, parS+pSB1C3, ori+pSB1K3, dif+pSB1A3)
· Transformation (Competent cell : E.coli DH10B)
· Spreading on plate (each antibiotics contained LB agar media)
8/3
Cell Culture
· Pick up a single colony from fresh cultured LB agar plate.
· Our established media and inoculate the cell into the 5ml of fresh LB liquid media (contains antibiotics) and our established media at 37℃ with shaking for 12hr.
· vector : parA+pSB1A3, parS+pSB1C3, ori+pSB1K3, dif+pSB1A3
8/4
Vector extract
· Uses kit and extracted plasmid.
(parA+pSB1A3, parS+pSB1C3, ori+pSB1K3, dif+pSB1A3)
· Confirmed size by electrophoresis.
8/9
Plasmid Digest
· Digest of 4 vector.
· ori+pSB1K3, parA+pSB1A3, parS+pSB1C3 digests by EcoRⅠ-SpeⅠ restriction endonuclease and dif+pSB1A3 gene digested by XbaⅠ-PstⅠ restriction endonuclease.
· Confirmed size by electrophoresis.
· Came out as size of ori+pSB1K3 gene is smaller.
8/10
Sequencing Order
· Analyzed plasmid DNA extraction (cultured cell)
8/16
Primer Again design
· According to analysis result, primer of ori gene knew wrong fact.
· Designed primer of ori gene again.
· Primer of I51020 and p1003 designed.
8/20
DNA Amplification
· Amplified I51020, p1003
· Confirmed size by electrophoresis.
8/23
Plasmid Digest
· I51020 digests by EcoRⅠ-PstⅠ restriction endonuclease.
· p1003 digests by XbaⅠ-PstⅠ restriction endonuclease.
· Confirmed size by electrophoresis.
8/24
DNA Ligation/Transformation
· Ligated inserting gene in vector
(Inserted ori gene and p1003 antibiotic resistance cassette in I51020 vector, inserted parS gene and dif gene in pSB1A3 vector)
· Transformation (Competent cell : E.coli DH10B)
· Spreading on plate (each antibiotics contained LB agar media)

8/25
I51020 transformation failed
Plan to design Cn vector
Ready to P1003, ori miniprep

9/2

Primer design for Cn vector. May be it won’t work….

All parts re-transformation for selection and miniprep.

 

9/7

Cn vector primer arrived.
Ready to PSB1C3 plasmid miniprep
Ori X/S digestion.

 

9/10

Cn vector PCR, purification, ligation.
Cn vector PCR Product electroporhesis.
9/15
Cn vector transformation failed.
Cn vercor PCR condition re arrangement
PrctB promoter primer design

 

9/25
Cn vector + ori(X-S) ligation, transformation.
PrctB primer arrived , PCR

9/27
Cn vector colony appear. But not red, may be failure.
PrctB + rctB ligation.
Cn vector transformation

9/30
Cn vector colony appear again. Not red too. Ready to miniprep.
parB + GFP fusion

10/3
parA, parS, dif ligation PSB1C3 and PSB1A3
computer engineering student meeting,

 

10/8
Ori PCR and miniprep
Ori digestion EP/XP/XS

 

10/10

parS, dif parts complete.
parA re PCR.

10/15
All members of CBNU-KOREA team meeting

10/21
Parts making start.

Ori function is not checked.

10/25

Parts sending
Prepare Team wiki freeze

Retrieved from "http://2010.igem.org/Team:CBNU-Korea/Notebook"