Team:Osaka/week1
From 2011.igem.org
(Difference between revisions)
(Created page with "==July 29 (Thu)== Attendance: Torigata, Takino, Teoh, Yasumoto, Kakuda, Saka, Tamura # Safety lecture for junior members. # Preparation of LB agar plates (26 Amp, 25 Kan, 25 Cam)...") |
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- | == | + | ==August 1 (Mon)== |
- | + | # Preparation of LB agar plates (33 Amp, 18 Kan, 17 Cam). | |
- | + | ||
- | # Preparation of LB agar plates ( | + | |
- | == | + | ==August 2 (Tue)== |
- | + | # Culture medium preparation | |
- | # | + | # Competent cells preparation - Nojima Method |
- | #* | + | #* SOB medium (MgCl2 not yet added) -> 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 (Wed)== | ||
+ | ==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]] | [[Team:Osaka/Notebook|Back to Notebook]] |
Revision as of 13:25, 11 August 2011
Contents |
August 1 (Mon)
- Preparation of LB agar plates (33 Amp, 18 Kan, 17 Cam).
August 2 (Tue)
- Culture medium preparation
- Competent cells preparation - Nojima Method
- SOB medium (MgCl2 not yet added) -> 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 (Wed)
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.
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
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 | (" ") |
1-5A | <bbpart>pSB1K3</bbpart> | K | (" ") |