Team:CBNU-Korea/Notebook

From 2010.igem.org

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==Notebook==
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5/7
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Membership meeting
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You should make use of the calendar feature on the wiki and start a lab notebook. This may be looked at by the judges to see how your work progressed throughout the summer. It is a very useful organizational tool as well.
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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
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Strategy about database
 +
Planned how to re-construct essential gene database.
 +
7/19
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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.
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• plasmid : pSB1C3, pSB1A3, pSB1T3, pSB1K3 (contain J54450)
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7/20
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Plasmid extract
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• Uses kit and extracted plasmid. (pSB1C3, pSB1A3, pSB1T3, pSB1K3)
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• Confirmed size by electrophoresis.
 +
7/23
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Plasmid Digest
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• Digest of 4 plasmid.
 +
• Used EcoRⅠ-PstⅠ restriction endonuclease.
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• Confirmed size by electrophoresis.
 +
7/24
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Primer Design
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• Designed Vibrio cholerae's oriC2vc, parA, parB, parS, dif gene primer using ‘Ginko primer’.
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7/28
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DNA Amplification
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• Amplified ori, parA, parB, parS, dif gene of vibrio cholerae.
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• Confirmed size by electrophoresis.
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Concentration
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• 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
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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
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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)

Revision as of 14:47, 27 October 2010

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Home Team Official Team Profile Project Parts Submitted to the Registry Modeling Notebook Safety


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. 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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