Team:CBNU-Korea/Notebook

From 2010.igem.org

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

Revision as of 18:43, 27 October 2010

<!DOCTYPE html PUBLIC "-//W3C//DTD XHTML 1.0 Transitional//EN" "http://www.w3.org/TR/xhtml1/DTD/xhtml1-transitional.dtd"> 무제 문서

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.

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.
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)

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