Team:Osaka/Notebook week2

From 2010.igem.org


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Week2 : August 1 - August 7

August 2 (Mon)

Attendance: Torigata, Takino, Teoh, Tadashi, Yasumoto, Saka

  1. Culture medium preparation
    • LB agar plates (49 antibiotic-less plates)
    • LB liquid medium (500 ml)
  2. 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

  1. 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

  1. OD measurements throughout the day till required OD (0.3~0.7) was obtained.
  2. Completion of competent cells according to protocol.

August 5 (Thu)

Attendance: Nakamura, Yasumoto, Saka, Kakuda, Takino

  1. Transformation of Registry parts:
IDPart NameResistanceDescription
2-20JBBa_K118023AC. fermi endocellulase Cen A coding
2-20HBBa_K118022AC. fermi exocellulase Cex coding
1-2MBBa_B0034ARBS
1-13DBBa_B0010Aterminator
1-1DBBa_R0010Apromoter
1-18FBBa_E1010KRFP coding

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.

August 6 (Fri)

Attendance: Nakamura, Saka, Yasumoto, Takino, Teoh

  1. 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
  2. Colonies transferred to 3ml LB growth medium & incubated o/n at 37˚C

August 7 (Sat)

Attendance: Nakamura, Saka, Yasumoto, Takino

  1. Miniprep of o/n-incubated colonies (2-20J, 2-20H, 1-2M, 1-13D, 1-1D, 1-18F) using Sigma-Aldrich Plasmid Miniprep Kit
  2. Transformation of construction plasmids
IDPart NameResistanceDescription
1-1CpSB1A3Aconstruction plasmid containing mRFP coding device (BBa_J04450)
1-3ApSB1C3C(" ")
1-5ApSB1K3K(" ")
  1. Meeting


July
Su Mo Tu We Th Fr Sa
1 2 3
4 5 6 7 8 9 10
11 12 13 14 15 16 17
18 19 20 21 22 23 24
Week1 25 26 27 28 29 30 31
August
Su Mo Tu We Th Fr Sa
Week2 1 2 3 4 5 6 7
Week3 8 9 10 11 12 13 14
Week4 15 16 17 18 19 20 21
Week5 22 23 24 25 26 27 28
Week6 29 30 31
September
Su Mo Tu We Th Fr Sa
Week6 1 2 3 4
Week7 5 6 7 8 9 10 11
Week8 12 13 14 15 16 17 18
Week9 19 20 21 22 23 24 25
Week10 26 27 28 29 30
October
Su Mo Tu We Th Fr Sa
Week10 1 2
Week11 3 4 5 6 7 8 9
Week12 10 11 12 13 14 15 16
Week13 17 18 19 20 21 22 23
Week14 24 25 26 27 28 29 30
31