Team:Osaka/week1

From 2011.igem.org

(Difference between revisions)
Line 1: Line 1:
==August 1 (Mon)==
==August 1 (Mon)==
# Preparation of LB agar plates (33 Amp, 18 Kan, 17 Cam).
# Preparation of LB agar plates (33 Amp, 18 Kan, 17 Cam).
 +
==August 2 (Tue)==
==August 2 (Tue)==
Line 6: Line 7:
# 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
 +
#* Preparation of glucose solution for making SOC medium.
#* 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.
-
==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).
# 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.
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.
Line 44: Line 30:
|}
|}
-
==August 6 (Fri)==
 
-
Attendance: Nakamura, Saka, Yasumoto, Takino, Teoh
 
-
# Colony check
 
-
#* All transformed cells produced colonies!
 
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#* Non-transformed cells (negative controls) did not grow on Amp, Kan or Cam plates -> confirmed lack of natural antibiotic resistance
 
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# Colonies transferred to 3ml LB growth medium & incubated o/n at 37˚C
 
-
==August 7 (Sat)==
+
==August 3 (Wed)==
-
Attendance: Nakamura, Saka, Yasumoto, Takino
+
# Competent cells preparation (continued)
-
# Miniprep of o/n-incubated colonies (2-20J, 2-20H, 1-2M, 1-13D, 1-1D, 1-18F) using Sigma-Aldrich Plasmid Miniprep Kit
+
#* Inoculation of pre-culture SOB from o/n-incubated agar plate colonies.
-
# Transformation of construction plasmids (See Table 2)
+
#* Transfer from pre-culture to growth culture.
-
# Meeting
+
 
 +
 
 +
==August 4 (Thu)==
 +
# OD measurements throughout the day till required OD (0.3~0.7) was obtained.
 +
# Completion of competent cells according to protocol.
 +
 
 +
 
 +
 
 +
==August 5 (Fri)==
 +
 
 +
 
 +
==August 6 (Sat)==
 +
# 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"
{|style="border-collapse: separate; border-spacing: 0; border-width: 1px; border-style: solid; border-color: #000; padding:5px;" cellpadding="5"
-
|+Table 2
+
|+Table 1
!ID!!Part Name!!Resistance!!Description
!ID!!Part Name!!Resistance!!Description
|-
|-
-
|1-1C||<bbpart>pSB1A3</bbpart>||A||construction plasmid containing mRFP coding device (<bbpart>BBa_J04450</bbpart>)
+
|2-20J||<bbpart>BBa_K118023</bbpart>||A||''C. fermi'' endocellulase Cen A coding
|-
|-
-
|1-3A||<bbpart>pSB1C3</bbpart>||C||<nowiki>(" ")</nowiki>
+
|2-20H||<bbpart>BBa_K118022</bbpart>||A||''C. fermi'' exocellulase Cex coding
|-
|-
-
|1-5A||<bbpart>pSB1K3</bbpart>||K||<nowiki>(" ")</nowiki>
+
|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
|}
|}

Revision as of 13:30, 11 August 2011

Contents

August 1 (Mon)

  1. Preparation of LB agar plates (33 Amp, 18 Kan, 17 Cam).


August 2 (Tue)

  1. Culture medium preparation
  2. Competent cells preparation - Nojima Method
    • SOB medium (MgCl2 not yet added) -> stored at 4˚C
    • Preparation of glucose solution for making SOC medium.
    • 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.

  1. 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.

Table 1
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


August 3 (Wed)

  1. Competent cells preparation (continued)
    • Inoculation of pre-culture SOB from o/n-incubated agar plate colonies.
    • Transfer from pre-culture to growth culture.


August 4 (Thu)

  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 (Fri)

August 6 (Sat)

  1. 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.

Table 1
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


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