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__NOTOC__ ==August 2 (Mon)== Attendance: Torigata, Takino, Teoh, Tadashi, Yasumoto, Saka # Culture medium preparation #* LB agar plates (49 antibiotic-less plates) #* LB liquid medium (500 ml) # Competent cells preparation - Nojima Method #* SOB medium (MgCl2 not yet added) -> stored at 4˚C #* TB buffer -> stored at 4˚C #* Single-colony streaking of E. coli (strain DH5α) on 2 LB agar plates -> 37˚C incubation o/n Tomorrow we shall prepare the remainder of the reagents and inoculate SOB with o/n-incubated E. coli. ==August 3 (Tue)== Attendance: Nakamura, Kakuda, Saka, Yasumoto, Teoh # Competent cells preparation (continued) #* Preparation of glucose solution for making SOC medium. #* Inoculation of pre-culture SOB from o/n-incubated agar plate colonies. #* (Night) Transfer from pre-culture to growth culture. ==August 4 (Wed)== Attendance: Nakamura, Saka, Kakuda, Yasumoto, Torigata, Teoh # OD measurements throughout the day till required OD (0.3~0.7) was obtained. # Completion of competent cells according to protocol. ==August 5 (Thu)== Attendance: Nakamura, Yasumoto, Saka, Kakuda, Takino # Transformation of Registry parts (See Table 1). 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. {|style="border-collapse: separate; border-spacing: 0; border-width: 1px; border-style: solid; border-color: #000; padding:5px;" cellpadding="5" |+Table 1 !ID!!Part Name!!Resistance!!Description |- |2-20J||<bbpart>BBa_K118023</bbpart>||A||''C. fermi'' endocellulase Cen A coding |- |2-20H||<bbpart>BBa_K118022</bbpart>||A||''C. fermi'' exocellulase Cex coding |- |1-2M||<bbpart>BBa_B0034</bbpart>||A||RBS |- |1-13D||<bbpart>BBa_B0010</bbpart>||A||terminator |- |1-1D||<bbpart>BBa_R0010</bbpart>||A||promoter |- |1-18F||<bbpart>BBa_E1010</bbpart>||K||RFP coding |} ==August 6 (Fri)== Attendance: Nakamura, Saka, Yasumoto, Takino, Teoh # Colony check #* All transformed cells produced colonies! #* Non-transformed cells (negative controls) did not grow on Amp, Kan or Cam plates -> confirmed lack of natural antibiotic resistance # Colonies transferred to 3ml LB growth medium & incubated o/n at 37˚C ==August 7 (Sat)== Attendance: Nakamura, Saka, Yasumoto, Takino # Miniprep of o/n-incubated colonies (2-20J, 2-20H, 1-2M, 1-13D, 1-1D, 1-18F) using Sigma-Aldrich Plasmid Miniprep Kit # Transformation of construction plasmids (See Table 2) # Meeting {|style="border-collapse: separate; border-spacing: 0; border-width: 1px; border-style: solid; border-color: #000; padding:5px;" cellpadding="5" |+Table 2 !ID!!Part Name!!Resistance!!Description |- |1-1C||<bbpart>pSB1A3</bbpart>||A||construction plasmid containing mRFP coding device (<bbpart>BBa_J04450</bbpart>) |- |1-3A||<bbpart>pSB1C3</bbpart>||C||<nowiki>(" ")</nowiki> |- |1-5A||<bbpart>pSB1K3</bbpart>||K||<nowiki>(" ")</nowiki> |} [[Team:Osaka/Notebook|Back to Notebook]]

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