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| - | {{Osaka}}
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| - | <div class="contents2">
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| - | {{Osaka/Calendar}}
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| - | <html>
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| - | <h2 class="margin">Week2:August 1 - August 7</h2>
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| - | <html><h3 id="0802">August 2 (Mon)</h3></html>
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| - | Attendance: Torigata, Takino, Teoh, Tadashi, Yasumoto, Saka
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| - | # Culture medium preparation
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| - | #* LB agar plates (49 antibiotic-less plates)
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| - | #* LB liquid medium (500 ml)
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| - | # Competent cells preparation - Nojima Method
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| - | #* SOB medium (MgCl2 not yet added) -> stored at 4˚C
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| - | #* TB buffer -> stored at 4˚C
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| - | #* Single-colony streaking of E. coli (strain DH5α) on 2 LB agar plates -> 37˚C incubation o/n
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| - | Tomorrow we shall prepare the remainder of the reagents and inoculate SOB with o/n-incubated E. coli.
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| - | <html><h3 id="0803">August 3 (Tue)</h3></html>
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| - | Attendance: Nakamura, Kakuda, Saka, Yasumoto, Teoh
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| - | # Competent cells preparation (continued)
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| - | #* Preparation of glucose solution for making SOC medium.
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| - | #* Inoculation of pre-culture SOB from o/n-incubated agar plate colonies.
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| - | #* (Night) Transfer from pre-culture to growth culture.
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| - | <html><h3 id="0804">August 4 (Wed)</h3></html>
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| - | Attendance: Nakamura, Saka, Kakuda, Yasumoto, Torigata, Teoh
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| - | # OD measurements throughout the day till required OD (0.3~0.7) was obtained.
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| - | # Completion of competent cells according to protocol.
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| - | <html><h3 id="0805">August 5 (Thu)</h3></html>
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| - | Attendance: Nakamura, Yasumoto, Saka, Kakuda, Takino
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| - | # Transformation of Registry parts:
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| - | {|
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| - | !ID!!Part Name!!Resistance!!Description
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| - | |-
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| - | |2-20J||<bbpart>BBa_K118023</bbpart>||A||''C. fermi'' endocellulase Cen A coding
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| - | |-
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| - | |2-20H||<bbpart>BBa_K118022</bbpart>||A||''C. fermi'' exocellulase Cex coding
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| - | |-
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| - | |1-2M||<bbpart>BBa_B0034</bbpart>||A||RBS
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| - | |-
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| - | |1-13D||<bbpart>BBa_B0010</bbpart>||A||terminator
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| - | |-
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| - | |1-1D||<bbpart>BBa_R0010</bbpart>||A||promoter
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| - | |-
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| - | |1-18F||<bbpart>BBa_E1010</bbpart>||K||RFP coding
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| - | |}
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| - | Note: 'ID' is an internal identifier used by wet work members to simplify labeling etc. In the case of iGEM distribution parts it usually refers to the plate location.
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| - | <html><h3 id="0806">August 6 (Fri)</h3></html>
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| - | Attendance: Nakamura, Saka, Yasumoto, Takino, Teoh
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| - | # Colony check
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| - | #* All transformed cells produced colonies!
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| - | #* Non-transformed cells (negative controls) did not grow on Amp, Kan or Cam plates -> confirmed lack of natural antibiotic resistance
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| - | # Colonies transferred to 3ml LB growth medium & incubated o/n at 37˚C
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| - | <html><h3 id="0807">August 7 (Sat)</h3></html>
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| - | Attendance: Nakamura, Saka, Yasumoto, Takino
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| - | # Miniprep of o/n-incubated colonies (2-20J, 2-20H, 1-2M, 1-13D, 1-1D, 1-18F) using Sigma-Aldrich Plasmid Miniprep Kit
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| - | # Transformation of construction plasmids
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| - | {|
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| - | !ID!!Part Name!!Resistance!!Description
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| - | |-
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| - | |1-1C||<bbpart>pSB1A3</bbpart>||A||construction plasmid containing mRFP coding device (<bbpart>BBa_J04450</bbpart>)
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| - | |-
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| - | |1-3A||<bbpart>pSB1C3</bbpart>||C||<nowiki>(" ")</nowiki>
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| - | |-
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| - | |1-5A||<bbpart>pSB1K3</bbpart>||K||<nowiki>(" ")</nowiki>
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| - | |}
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| - | # Meeting
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| - | <html><hr class="margin">
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| - | </html>
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| - | {{Osaka/Calendar}}
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| - | <html>
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| - | </div>
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| - | </html>
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