Team:Kyoto/Notebook

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{{:Team:Kyoto/Header}}
{{:Team:Kyoto/Header}}
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==Index==
 
==Notebook==
==Notebook==
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===Tuesday, July 20 <span class="by">By: Wataru, Tomo, Yuki, Kazuya, Ken, Makoto</span>===
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===Notebooks===
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<div class="preparation">
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* [[Team:Kyoto/Notebook1|Notebook1]]: Construction for Lysisbox.
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====Solubilization of Antibiotics====
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* [[Team:Kyoto/Notebook2|Notebook2]]: Measurement of R0011.
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{| class="experiments"
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* [[Team:Kyoto/Notebook3|Notebook3]]: Measurement of Lysis Cassette and Lysisbox etc.
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|Ampicillin(Amp)||Kanamycin(Kan)||
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|-
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[[#top-section|^Top]]
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|Mix 1.0g Amp and 20ml MilliQ||Mix 0.5g Kan and 10ml MilliQ||Final concentration is 50mg/ml
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|-
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===Other Information===
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|colspan="2"|Dispense 1.1ml of the solution into 1.5ml tubes||
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* [[Team:Kyoto/Protocols|Protocols]]: Protocols of each experiment such as Polymerase Chain Reaction (PCR), Restriction Digestion, Ligation, Transformation.
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|-
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* [[Team:Kyoto/Materials|Materials]]: Strains, DNA, and Primers.
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|colspan="2"|Store in the freezer (-20&#x2103;)||
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* [[Team:Kyoto/Parts|Parts]]: Construction of each part and BioBrick Parts used in our project.
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|}
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</div>
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[[#top-section|^Top]]
 +
 
----
----
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<div class="plate">
 
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====Making plates for LB (Amp+) and LB (Kan+)====
 
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</div>
 
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----
 
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<div class="transformation lysis measure">
 
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====[[Team:Kyoto/Protocols#Transformation|Transformation]]====
 
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{| class="experiments"
 
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!Name||Well||Sample (&micro;l)||Competent Cells (&micro;l)||Total (&micro;l)||Plate||Incubation||Result
 
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|-
 
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|<partinfo>J23100</partinfo>||1-18-C||1||20||21||rowspan="7"|LB (Amp+)||rowspan="8"|At 37&#x2103;, 7/20 20:50 - 7/21 17:00||&#x25CB;
 
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|-
 
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|<partinfo>J23105</partinfo>||1-18-M||1||20||21||&#x25CB;
 
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|-
 
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|<partinfo>J23116</partinfo>||1-20-M||1||20||21||&#x25CB;
 
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|-
 
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|<partinfo>R0011</partinfo>||1-6-G||1||20||21||&#x25CB;
 
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|-
 
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|<partinfo>E0840</partinfo>||1-12-O||1||20||21||&#x25CB;
 
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|-
 
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|<partinfo>J06702</partinfo>||2-8-E||1||20||21||&#x25CB;
 
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|-
 
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|<partinfo>pSB4K5</partinfo>||1-5-G||1||20||21||&#x25CF;
 
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|-
 
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|<partinfo>B0015</partinfo>||1-23-L||1||20||21||LB (Kan+)||&#x25CF;
 
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|}
 
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A vector of "pSB4K5" is Kanamycin-resistance, however, we plated it to LB plate (Amp+). And We didn't pre-culture "B0015" despite its vector is Kanamycin-resistance. So, it was predicted that we will fail the transformation of "pSB4K5" and "B0015".
 
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</div>
 
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===Wednesday, July 21 <span class="by">By: Wataru, Ken, Makoto, Takuya Y.</span>===
 
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<div class="culture lysis measure">
 
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====Culture of plates in which colonies was observed at 37&#x2103; from 07/21 20:50 to 07/22 17:00====
 
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</div>
 
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----
 
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<div class="master lysis measure">
 
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====Making a master plate of the above plates====
 
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</div>
 
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----
 
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<div class="transformation lysis measure">
 
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====Retry [[Team:Kyoto/Protocols#Transformation|Transformation]]====
 
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{| class="experiments"
 
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!Name||Well||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 (Kan+)||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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</div>
 
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----
 
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<div class="pcr lysis">
 
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====[[Team:Kyoto/Protocols#Stantard_PCR|PCR]] for S-R-Rz/Rz1 and S====
 
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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 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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{|class="experiments"
 
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|+ PCR condition
 
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|-
 
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|94&#x2103;||2min||
 
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|-
 
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|98&#x2103;||10sec||rowspan="3"|30 cycles
 
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|-
 
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|55&#x2103;||30sec
 
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|-
 
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|68&#x2103;||4min
 
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|-
 
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|4&#x2103;||forever||
 
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|}
 
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</div>
 

Latest revision as of 11:33, 27 October 2010

Contents

Notebook

Notebooks

^Top

Other Information

  • Protocols: Protocols of each experiment such as Polymerase Chain Reaction (PCR), Restriction Digestion, Ligation, Transformation.
  • Materials: Strains, DNA, and Primers.
  • Parts: Construction of each part and BioBrick Parts used in our project.

^Top

Retrieved from "http://2010.igem.org/Team:Kyoto/Notebook"