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| | {{:Team:Kyoto/Header}} | | {{:Team:Kyoto/Header}} |
| - | ==Index==
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| - |
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| | ==Notebook== | | ==Notebook== |
| | + | ===Notebooks=== |
| | + | * [[Team:Kyoto/Notebook1|Notebook1]]: Construction for Lysisbox. |
| | + | * [[Team:Kyoto/Notebook2|Notebook2]]: Measurement of R0011. |
| | + | * [[Team:Kyoto/Notebook3|Notebook3]]: Measurement of Lysis Cassette and Lysisbox etc. |
| | + | |
| | + | [[#top-section|^Top]] |
| | + | |
| | + | ===Other Information=== |
| | + | * [[Team:Kyoto/Protocols|Protocols]]: Protocols of each experiment such as Polymerase Chain Reaction (PCR), Restriction Digestion, Ligation, Transformation. |
| | + | * [[Team:Kyoto/Materials|Materials]]: Strains, DNA, and Primers. |
| | + | * [[Team:Kyoto/Parts|Parts]]: Construction of each part and BioBrick Parts used in our project. |
| | + | |
| | + | [[#top-section|^Top]] |
| | | | |
| - | {| class="note"
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| - | |+Tuesday, July 20
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| - | |-
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| - | |
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| - | *By: Wataru, Tomo, Yuki, Kazuya, Ken, Makoto
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| - | *Category: Antibiotic, Transformation
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| - | |-
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| - | |[[Team:Kyoto/Protocols#Solubilization_of_Antibiotics|Solubilized of Antibiotics]], Ampicillin (1g) and Kanamycin (0.5g).
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| - | |-
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| - | |[[Team:Kyoto/Protocols#Making_Plate|Made plates]] for LB (Ampicillin+) and LB (Kanamycin+).
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| - | |-
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| - | |[[Team:Kyoto/Protocols#Transformation|Transformed]] iGEM Parts.
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| - | {| class="experiments"
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| - | !Name||Well<sup>*1</sup>||Sample (µl)||Competent Cells (µl)||Total (µl)||Plate||Incubation||Result
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| - | |-
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| - | |<partinfo>J23100</partinfo>||1-18-C||1||20||21||rowspan="7"|LB (Ampicillin+)||rowspan="8"|At 37℃ 7/20 20:50 - 7/21 17:00||○
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| - | |-
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| - | |<partinfo>J23105</partinfo>||1-18-M||1||20||21||○
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| - | |-
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| - | |<partinfo>J23116</partinfo>||1-20-M||1||20||21||○
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| - | |-
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| - | |<partinfo>R0011</partinfo>||1-6-G||1||20||21||○
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| - | |-
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| - | |<partinfo>E0840</partinfo>||1-12-O||1||20||21||○
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| - | |-
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| - | |<partinfo>J06702</partinfo>||2-8-E||1||20||21||○
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| - | |-
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| - | |<partinfo>pSB4K5</partinfo>||1-5-G||1||20||21||×
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| - | |-
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| - | |<partinfo>B0015</partinfo>||1-23-L||1||20||21||LB (Kanamycin+)||×
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| - | |}
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| - | * *1 "1-18-C" means well 18C in [http://partsregistry.org/Help:Spring_2010_DNA_distribution Spring 2010 DNA Distribution Kit] Plate 1.
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| - | * '''Discussion'''
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| - | * A vector of "pSB4K5" is Kanamycin-resistance, however, we plated it to LB plate (Ampicillin+).
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| - | * And We didn't pre-culture "B0015" despite its vector is Kanamycin-resistance.
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| - | * So, it was predicted that we will fail the transformation of "pSB4K5" and "B0015".
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| - | |}
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| - | ----
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| - | {| class="note"
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| - | |+Wednesday, July 21
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| - | |-
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| - | |
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| - | *By: Wataru, Ken, Makoto, Takuya Yamamoto
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| - | *Category: Transformation, PCR, Lysis Cassette
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| - | |-
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| - | |Cultured plates in which colonies was observed at 37℃ from 07/21 20:50 to 07/22 17:00.
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| - | |-
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| - | |Made a master plate of the above plates.
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| - | |-
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| - | |Retried Transformation of iGEM Parts.
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| - | {| class="experiments"
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| - | |Name||Well<sup>*1</sup>||Sample (µl)||Competent Cells (µl)||Total (µl)||Plate||Incubation||Result
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| - | |-
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| - | |<partinfo>pSB4K5</partinfo>||1-5-G||1||20||21||rowspan="2"|LB (Kanamycin+)||rowspan="2"|At 37℃ 7/21 20:50 - 7/22 16:30||○
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| - | |-
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| - | |<partinfo>B0015</partinfo>||1-23-L||1||20||21||○
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| - | |}
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| - | * *1 "1-5-G" means well 5G in [http://partsregistry.org/Help:Spring_2010_DNA_distribution Spring 2010 DNA Distribution Kit] Plate 1.
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| - | |-
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| - | |PCR PCR for S-R-Rz/Rz1 and S
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| - | * Dilute λDNA (0.5µg/µl) 100 times with MilliQ. The final concentration of template λDNA was 5ng/µl.
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| - | {| class="experiments"
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| - | !No.||Water||25mM MgSO4||2mM dNTPs||10xBuffer for KOD Plus ver.2||TemplateDNA (5ng/µl)||Primer S-R-Rz/Rz1 Forward (10µM)||Primer S-R-Rz/Rz1 Reverse (10µM)||Primer S Reverse (10µM)||KOD Plus ver.2||Total
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| - | |-
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| - | |1||28µl||3µl||5µl||5µl||5µl||1.5µl||1.5µl||-||1µl||50µl
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| - | |-
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| - | |2||28µl||3µl||5µl||5µl||5µl||1.5µl||1.5µl||-||1µl||50µl
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| - | |-
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| - | |3||28µl||3µl||5µl||5µl||5µl||1.5µl||-||1.5µl||1µl||50µl
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| - | |-
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| - | |4||28µl||3µl||5µl||5µl||5µl||1.5µl||-||1.5µl||1µl||50µl
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| - | |-
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| - | |5||28µl||3µl||5µl||5µl||5µl||1.5µl||1.5µl||-||1µl||50µl
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| - | |-
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| - | |6||28µl||3µl||5µl||5µl||5µl||1.5µl||1.5µl||-||1µl||50µl
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| - | |-
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| - | |7||28µl||3µl||5µl||5µl||5µl||1.5µl||-||1.5µl||1µl||50µl
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| - | |-
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| - | |8||28µl||3µl||5µl||5µl||5µl||1.5µl||-||1.5µl||1µl||50µl
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| - | |}
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| - | * Forward Primer of S-R-Rz/Rz1 and S is common.
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| - | * PCR condition : 94℃ x 2min, (98℃ x 10sec, 55℃ x 30sec, 68℃ x 1min) x 30cycles, 4℃ forever
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| - | |}
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| | ---- | | ---- |
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