Team:Osaka/Notebook

From 2010.igem.org

(Difference between revisions)
(→August 9 (Mon))
(→Notebook)
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!ID!!Part Name!!Resistance!!Description
!ID!!Part Name!!Resistance!!Description
|-
|-
-
|2-20J||<partinfo>BBa_K118023</partinfo>||A||''C. fermi'' endocellulase Cen A coding
+
|2-20J||<bbpart>BBa_K118023</bbpart>||A||''C. fermi'' endocellulase Cen A coding
|-
|-
-
|2-20H||<partinfo>BBa_K118022</partinfo>||A||''C. fermi'' exocellulase Cex coding
+
|2-20H||<bbpart>BBa_K118022</bbpart>||A||''C. fermi'' exocellulase Cex coding
|-
|-
-
|1-2M||<partinfo>BBa_B0034</partinfo>||A||RBS
+
|1-2M||<bbpart>BBa_B0034</bbpart>||A||RBS
|-
|-
-
|1-13D||<partinfo>BBa_B0010</partinfo>||A||terminator
+
|1-13D||<bbpart>BBa_B0010</bbpart>||A||terminator
|-
|-
-
|1-1D||<partinfo>BBa_R0010</partinfo>||A||promoter
+
|1-1D||<bbpart>BBa_R0010</bbpart>||A||promoter
|-
|-
-
|1-18F||<partinfo>BBa_E1010</partinfo>||K||RFP coding
+
|1-18F||<bbpart>BBa_E1010</bbpart>||K||RFP 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.
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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!ID!!Part Name!!Resistance!!Description
!ID!!Part Name!!Resistance!!Description
|-
|-
-
|1-1C||<partinfo>pSB1A3</partinfo>||A||construction plasmid containing mRFP coding device (<partinfo>BBa_J04450</partinfo>)
+
|1-1C||<bbpart>pSB1A3</bbpart>||A||construction plasmid containing mRFP coding device (<bbpart>BBa_J04450</bbpart>)
|-
|-
-
|1-3A||<partinfo>pSB1C3</partinfo>||C||<nowiki>(" ")</nowiki>
+
|1-3A||<bbpart>pSB1C3</bbpart>||C||<nowiki>(" ")</nowiki>
|-
|-
-
|1-5A||<partinfo>pSB1K3</partinfo>||K||<nowiki>(" ")</nowiki>
+
|1-5A||<bbpart>pSB1K3</bbpart>||K||<nowiki>(" ")</nowiki>
|}
|}
# Meeting
# Meeting
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# Transfer of 1-3A, 1-5A colonies to solution culture (repeat)
# Transfer of 1-3A, 1-5A colonies to solution culture (repeat)
#*''Yesterday's inoculated culture mediums contained the wrong antibiotics!''
#*''Yesterday's inoculated culture mediums contained the wrong antibiotics!''
 +
# Transformation of <bbpart>BBa_I13521</bbpart>, <bbpart>BBa_I13522</bbpart>, <bbpart>BBa_I13600</bbpart>, <bbpart>BBa_K204031</bbpart>, <bbpart>BBa_K204051</bbpart>, <bbpart>BBa_K204032</bbpart> from last year's stock
 +
 +
===August 10 (Tue)===
 +
# Miniprep of 1-3A, 1-5A
 +
# Restriction digests of 1-3A, 1-5A
 +
# Gel electrophoresis (twice)
 +
 +
===August 11 (Wed)===
 +
# Miniprep of last year's parts transformed on Monday
 +
# Gel electrophoresis of 1-1D, 1-18F, 1-13D, 1-2M
 +
#* all 4 plasmids could not be cut... AGAIN
 +
#* so far all restriction digests involving XbaI seem to have failed; problem with our enzyme?
 +
#* will try with different set of restriction enzymes next week
 +
 +
===August 16 (Mon)===

Revision as of 14:48, 16 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<bbpart>BBa_K118023</bbpart>AC. fermi endocellulase Cen A coding
2-20H<bbpart>BBa_K118022</bbpart>AC. fermi exocellulase Cex coding
1-2M<bbpart>BBa_B0034</bbpart>ARBS
1-13D<bbpart>BBa_B0010</bbpart>Aterminator
1-1D<bbpart>BBa_R0010</bbpart>Apromoter
1-18F<bbpart>BBa_E1010</bbpart>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<bbpart>pSB1A3</bbpart>Aconstruction plasmid containing mRFP coding device (<bbpart>BBa_J04450</bbpart>)
1-3A<bbpart>pSB1C3</bbpart>C(" ")
1-5A<bbpart>pSB1K3</bbpart>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!
  4. Transformation of <bbpart>BBa_I13521</bbpart>, <bbpart>BBa_I13522</bbpart>, <bbpart>BBa_I13600</bbpart>, <bbpart>BBa_K204031</bbpart>, <bbpart>BBa_K204051</bbpart>, <bbpart>BBa_K204032</bbpart> from last year's stock

August 10 (Tue)

  1. Miniprep of 1-3A, 1-5A
  2. Restriction digests of 1-3A, 1-5A
  3. Gel electrophoresis (twice)

August 11 (Wed)

  1. Miniprep of last year's parts transformed on Monday
  2. Gel electrophoresis of 1-1D, 1-18F, 1-13D, 1-2M
    • all 4 plasmids could not be cut... AGAIN
    • so far all restriction digests involving XbaI seem to have failed; problem with our enzyme?
    • will try with different set of restriction enzymes next week

August 16 (Mon)

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