Team:Osaka/Notebook

From 2010.igem.org

(Difference between revisions)
(→Notebook)
(→Notebook)
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Attendance: Miyatake, Hirayama, Torigata, Teoh, Tadashi, Yasumoto, Kakuda, Saka
Attendance: Miyatake, Hirayama, Torigata, Teoh, Tadashi, Yasumoto, Kakuda, Saka
# Meeting
# Meeting
-
## Summer project schedule
+
#* Summer project schedule
-
## List of genes to clone
+
#* List of genes to clone
===August 2 (Mon)===
===August 2 (Mon)===
Attendance: Torigata, Takino, Teoh, Tadashi, Yasumoto, Saka
Attendance: Torigata, Takino, Teoh, Tadashi, Yasumoto, Saka
# Culture medium preparation
# Culture medium preparation
-
## LB agar plates (49 antibiotic-less plates)
+
#* LB agar plates (49 antibiotic-less plates)
-
## LB liquid medium (500 ml)
+
#* LB liquid medium (500 ml)
# Competent cells preparation - Nojima Method
# Competent cells preparation - Nojima Method
-
## SOB medium (MgCl2 not yet added) -> stored at 4˚C
+
#* SOB medium (MgCl2 not yet added) -> stored at 4˚C
-
## TB buffer -> 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
+
#* 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.
Tomorrow we shall prepare the remainder of the reagents and inoculate SOB with o/n-incubated E. coli.
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Attendance: Nakamura, Kakuda, Saka, Yasumoto, Teoh
Attendance: Nakamura, Kakuda, Saka, Yasumoto, Teoh
# Competent cells preparation (continued)
# Competent cells preparation (continued)
-
## Preparation of glucose solution for making SOC medium.
+
#* Preparation of glucose solution for making SOC medium.
-
## Inoculation of pre-culture SOB from o/n-incubated agar plate colonies.
+
#* Inoculation of pre-culture SOB from o/n-incubated agar plate colonies.
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## (Night) Transfer from pre-culture to growth culture.
+
#* (Night) Transfer from pre-culture to growth culture.
===August 4 (Wed)===
===August 4 (Wed)===
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Attendance: Nakamura, Saka, Yasumoto, Takino, Teoh
Attendance: Nakamura, Saka, Yasumoto, Takino, Teoh
# Colony check
# Colony check
-
## All transformed cells produced colonies!
+
#* 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
+
#* Non-transformed cells (negative controls) did not grow on Amp, Kan or Cam plates -> confirmed lack of natural antibiotic resistance
# Colonies transferred to LB growth medium & incubated o/n at 37˚C
# Colonies transferred to LB growth medium & incubated o/n at 37˚C

Revision as of 04:10, 9 August 2010

Home Team Project Submitted Parts Modeling Protocols Notebook Safety


Contents

Notebook

July 29 (Thu)

Attendance: Torigata, Takino, Teoh, Yasumoto, Kakuda, Saka, Tamura

  1. Safety lecture for junior members.
  2. Preparation of LB agar plates (26 Amp, 25 Kan, 25 Cam).

July 31 (Thu)

Attendance: Miyatake, Hirayama, Torigata, Teoh, Tadashi, Yasumoto, Kakuda, Saka

  1. Meeting
    • Summer project schedule
    • List of genes to clone

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
2-20J<partinfo>BBa_K118023</partinfo>AC. fermi endocellulase Cen A coding
2-20H<partinfo>BBa_K118022</partinfo>AC. fermi exocellulase Cex coding
1-2M<partinfo>BBa_B0034</partinfo>ARBS
1-13D<partinfo>BBa_B0010</partinfo>Aterminator
1-1D<partinfo>BBa_R0010</partinfo>Apromoter
1-18F<partinfo>BBa_E1010</partinfo>KRFP 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 LB growth medium & incubated o/n at 37˚C

August 7 (Sat)

Attendance: Nakamura, Saka, Yasumoto, Takino

  1. Miniprep
  2. Transformation of construction plasmids
IDPart NameResistanceDescription
1-1C<partinfo>pSB1A3</partinfo>Aconstruction plasmid containing mRFP coding device (<partinfo>BBa_J04450</partinfo>)
1-3A<partinfo>pSB1C3</partinfo>C(" ")
1-5A<partinfo>pSB1K3</partinfo>K(" ")
  1. Meeting

August 8 (Sun)

Attendance: Nakamura, Yasumoto

  1. Colony check
    All parts successfully transformed
  2. Transfer to liquid culture medium

August 9 (Mon)

  1. Miniprep of 1-1C
  2. Restriction digests of 2-20H, 2-20J, 1-1D, 1-18F, 1-1C, 1-13D, 1-2M
  3. Transfer of 1-3A, 1-5A colonies to solution culture (repeat)
    Yesterday's inoculated culture mediums contained the wrong antibiotics!
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