"

From 2010.igem.org

Home	Team	Project	Submitted Parts	Modeling	Notebook	Safety

Contents 1 Notebook 1.1 July 29 (Thu) 1.2 July 31 (Thu) 1.3 August 2 (Mon) 1.4 August 3 (Tue) 1.5 August 4 (Wed) 1.6 August 5 (Thu) 1.7 August 6 (Fri) 1.8 August 7 (Sat) 1.9 August 8 (Sun) 1.10 August 9 (Mon)

Notebook

August M T W T F S S             [http://2010.igem.org/wiki/index.php?title=August/1_August_2010&action=edit 1] [http://2010.igem.org/August/2_August_2010 2] [http://2010.igem.org/wiki/index.php?title=August/3_August_2010&action=edit 3] [http://2010.igem.org/wiki/index.php?title=August/4_August_2010&action=edit 4] [http://2010.igem.org/wiki/index.php?title=August/5_August_2010&action=edit 5] [http://2010.igem.org/wiki/index.php?title=August/6_August_2010&action=edit 6] [http://2010.igem.org/wiki/index.php?title=August/7_August_2010&action=edit 7] [http://2010.igem.org/wiki/index.php?title=August/8_August_2010&action=edit 8] [http://2010.igem.org/wiki/index.php?title=August/9_August_2010&action=edit 9] [http://2010.igem.org/wiki/index.php?title=August/10_August_2010&action=edit 10] [http://2010.igem.org/wiki/index.php?title=August/11_August_2010&action=edit 11] [http://2010.igem.org/wiki/index.php?title=August/12_August_2010&action=edit 12] [http://2010.igem.org/wiki/index.php?title=August/13_August_2010&action=edit 13] [http://2010.igem.org/wiki/index.php?title=August/14_August_2010&action=edit 14] [http://2010.igem.org/wiki/index.php?title=August/15_August_2010&action=edit 15] [http://2010.igem.org/wiki/index.php?title=August/16_August_2010&action=edit 16] [http://2010.igem.org/wiki/index.php?title=August/17_August_2010&action=edit 17] [http://2010.igem.org/wiki/index.php?title=August/18_August_2010&action=edit 18] [http://2010.igem.org/wiki/index.php?title=August/19_August_2010&action=edit 19] [http://2010.igem.org/wiki/index.php?title=August/20_August_2010&action=edit 20] [http://2010.igem.org/wiki/index.php?title=August/21_August_2010&action=edit 21] [http://2010.igem.org/wiki/index.php?title=August/22_August_2010&action=edit 22] [http://2010.igem.org/wiki/index.php?title=August/23_August_2010&action=edit 23] [http://2010.igem.org/wiki/index.php?title=August/24_August_2010&action=edit 24] [http://2010.igem.org/wiki/index.php?title=August/25_August_2010&action=edit 25] [http://2010.igem.org/wiki/index.php?title=August/26_August_2010&action=edit 26] [http://2010.igem.org/wiki/index.php?title=August/27_August_2010&action=edit 27] [http://2010.igem.org/wiki/index.php?title=August/28_August_2010&action=edit 28] [http://2010.igem.org/wiki/index.php?title=August/29_August_2010&action=edit 29] [http://2010.igem.org/wiki/index.php?title=August/30_August_2010&action=edit 30] [http://2010.igem.org/wiki/index.php?title=August/31_August_2010&action=edit 31]

September M T W T F S S     [http://2010.igem.org/wiki/index.php?title=September/1_September_2010&action=edit 1] [http://2010.igem.org/wiki/index.php?title=September/2_September_2010&action=edit 2] [http://2010.igem.org/wiki/index.php?title=September/3_September_2010&action=edit 3] [http://2010.igem.org/wiki/index.php?title=September/4_September_2010&action=edit 4] [http://2010.igem.org/wiki/index.php?title=September/5_September_2010&action=edit 5] [http://2010.igem.org/wiki/index.php?title=September/6_September_2010&action=edit 6] [http://2010.igem.org/wiki/index.php?title=September/7_September_2010&action=edit 7] [http://2010.igem.org/wiki/index.php?title=September/8_September_2010&action=edit 8] [http://2010.igem.org/wiki/index.php?title=September/9_September_2010&action=edit 9] [http://2010.igem.org/wiki/index.php?title=September/10_September_2010&action=edit 10] [http://2010.igem.org/wiki/index.php?title=September/11_September_2010&action=edit 11] [http://2010.igem.org/wiki/index.php?title=September/12_September_2010&action=edit 12] [http://2010.igem.org/wiki/index.php?title=September/13_September_2010&action=edit 13] [http://2010.igem.org/wiki/index.php?title=September/14_September_2010&action=edit 14] [http://2010.igem.org/wiki/index.php?title=September/15_September_2010&action=edit 15] [http://2010.igem.org/wiki/index.php?title=September/16_September_2010&action=edit 16] [http://2010.igem.org/wiki/index.php?title=September/17_September_2010&action=edit 17] [http://2010.igem.org/wiki/index.php?title=September/18_September_2010&action=edit 18] [http://2010.igem.org/wiki/index.php?title=September/19_September_2010&action=edit 19] [http://2010.igem.org/wiki/index.php?title=September/20_September_2010&action=edit 20] [http://2010.igem.org/wiki/index.php?title=September/21_September_2010&action=edit 21] [http://2010.igem.org/wiki/index.php?title=September/22_September_2010&action=edit 22] [http://2010.igem.org/wiki/index.php?title=September/23_September_2010&action=edit 23] [http://2010.igem.org/wiki/index.php?title=September/24_September_2010&action=edit 24] [http://2010.igem.org/wiki/index.php?title=September/25_September_2010&action=edit 25] [http://2010.igem.org/wiki/index.php?title=September/26_September_2010&action=edit 26] [http://2010.igem.org/wiki/index.php?title=September/27_September_2010&action=edit 27] [http://2010.igem.org/wiki/index.php?title=September/28_September_2010&action=edit 28] [http://2010.igem.org/wiki/index.php?title=September/29_September_2010&action=edit 29] [http://2010.igem.org/wiki/index.php?title=September/30_September_2010&action=edit 30]

October M T W T F S S         [http://2010.igem.org/wiki/index.php?title=October/1_October_2010&action=edit 1] [http://2010.igem.org/wiki/index.php?title=October/2_October_2010&action=edit 2] [http://2010.igem.org/wiki/index.php?title=October/3_October_2010&action=edit 3] [http://2010.igem.org/wiki/index.php?title=October/4_October_2010&action=edit 4] [http://2010.igem.org/wiki/index.php?title=October/5_October_2010&action=edit 5] [http://2010.igem.org/wiki/index.php?title=October/6_October_2010&action=edit 6] [http://2010.igem.org/wiki/index.php?title=October/7_October_2010&action=edit 7] [http://2010.igem.org/wiki/index.php?title=October/8_October_2010&action=edit 8] [http://2010.igem.org/wiki/index.php?title=October/9_October_2010&action=edit 9] [http://2010.igem.org/wiki/index.php?title=October/10_October_2010&action=edit 10] [http://2010.igem.org/wiki/index.php?title=October/11_October_2010&action=edit 11] [http://2010.igem.org/wiki/index.php?title=October/12_October_2010&action=edit 12] [http://2010.igem.org/wiki/index.php?title=October/13_October_2010&action=edit 13] [http://2010.igem.org/wiki/index.php?title=October/14_October_2010&action=edit 14] [http://2010.igem.org/wiki/index.php?title=October/15_October_2010&action=edit 15] [http://2010.igem.org/wiki/index.php?title=October/16_October_2010&action=edit 16] [http://2010.igem.org/wiki/index.php?title=October/17_October_2010&action=edit 17] [http://2010.igem.org/wiki/index.php?title=October/18_October_2010&action=edit 18] [http://2010.igem.org/wiki/index.php?title=October/19_October_2010&action=edit 19] [http://2010.igem.org/wiki/index.php?title=October/20_October_2010&action=edit 20] [http://2010.igem.org/wiki/index.php?title=October/21_October_2010&action=edit 21] [http://2010.igem.org/wiki/index.php?title=October/22_October_2010&action=edit 22] [http://2010.igem.org/wiki/index.php?title=October/23_October_2010&action=edit 23] [http://2010.igem.org/wiki/index.php?title=October/24_October_2010&action=edit 24] [http://2010.igem.org/wiki/index.php?title=October/25_October_2010&action=edit 25] [http://2010.igem.org/wiki/index.php?title=October/26_October_2010&action=edit 26] [http://2010.igem.org/wiki/index.php?title=October/27_October_2010&action=edit 27] [http://2010.igem.org/wiki/index.php?title=October/28_October_2010&action=edit 28] [http://2010.igem.org/wiki/index.php?title=October/29_October_2010&action=edit 29] [http://2010.igem.org/wiki/index.php?title=October/30_October_2010&action=edit 30] [http://2010.igem.org/wiki/index.php?title=October/31_October_2010&action=edit 31]

November M T W T F S S [http://2010.igem.org/wiki/index.php?title=November/1_November_2010&action=edit 1] [http://2010.igem.org/wiki/index.php?title=November/2_November_2010&action=edit 2] [http://2010.igem.org/wiki/index.php?title=November/3_November_2010&action=edit 3] [http://2010.igem.org/wiki/index.php?title=November/4_November_2010&action=edit 4] [http://2010.igem.org/wiki/index.php?title=November/5_November_2010&action=edit 5] [http://2010.igem.org/wiki/index.php?title=November/6_November_2010&action=edit 6] [http://2010.igem.org/wiki/index.php?title=November/7_November_2010&action=edit 7] [http://2010.igem.org/wiki/index.php?title=November/8_November_2010&action=edit 8] [http://2010.igem.org/wiki/index.php?title=November/9_November_2010&action=edit 9] [http://2010.igem.org/wiki/index.php?title=November/10_November_2010&action=edit 10] [http://2010.igem.org/wiki/index.php?title=November/11_November_2010&action=edit 11] [http://2010.igem.org/wiki/index.php?title=November/12_November_2010&action=edit 12] [http://2010.igem.org/wiki/index.php?title=November/13_November_2010&action=edit 13] [http://2010.igem.org/wiki/index.php?title=November/14_November_2010&action=edit 14] [http://2010.igem.org/wiki/index.php?title=November/15_November_2010&action=edit 15] [http://2010.igem.org/wiki/index.php?title=November/16_November_2010&action=edit 16] [http://2010.igem.org/wiki/index.php?title=November/17_November_2010&action=edit 17] [http://2010.igem.org/wiki/index.php?title=November/18_November_2010&action=edit 18] [http://2010.igem.org/wiki/index.php?title=November/19_November_2010&action=edit 19] [http://2010.igem.org/wiki/index.php?title=November/20_November_2010&action=edit 20] [http://2010.igem.org/wiki/index.php?title=November/21_November_2010&action=edit 21] [http://2010.igem.org/wiki/index.php?title=November/22_November_2010&action=edit 22] [http://2010.igem.org/wiki/index.php?title=November/23_November_2010&action=edit 23] [http://2010.igem.org/wiki/index.php?title=November/24_November_2010&action=edit 24] [http://2010.igem.org/wiki/index.php?title=November/25_November_2010&action=edit 25] [http://2010.igem.org/wiki/index.php?title=November/26_November_2010&action=edit 26] [http://2010.igem.org/wiki/index.php?title=November/27_November_2010&action=edit 27] [http://2010.igem.org/wiki/index.php?title=November/28_November_2010&action=edit 28] [http://2010.igem.org/wiki/index.php?title=November/29_November_2010&action=edit 29] [http://2010.igem.org/wiki/index.php?title=November/30_November_2010&action=edit 30]

Note: For detailed lab notes see individual pages in calendar above.

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
   1. Summer project schedule
   2. List of genes to clone

August 2 (Mon)

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

1. Culture medium preparation
   1. LB agar plates (49 antibiotic-less plates)
   2. LB liquid medium (500 ml)
2. Competent cells preparation - Nojima Method
   1. SOB medium (MgCl2 not yet added) -> stored at 4˚C
   2. TB buffer -> stored at 4˚C
   3. 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)
   1. Preparation of glucose solution for making SOC medium.
   2. Inoculation of pre-culture SOB from o/n-incubated agar plate colonies.
   3. (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 basic Registry parts:

ID	Part Name	Resistance	Description
2-20J	BBa_K118023	A	C. fermi endocellulase Cen A coding
2-20H	BBa_K118022	A	C. fermi exocellulase Cex coding
1-2M	BBa_B0034	A	RBS
1-13D	BBa_B0010	A	Terminator
1-1D	BBa_R0010	A	Promoter
1-18F	BBa_E1010	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.

August 6 (Fri)

1. Colony check
   1. All transformed cells produced colonies!
   2. 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)

1. Miniprep
2. Transformation
3. Meeting

August 8 (Sun)

1. Transfer to liquid culture medium

August 9 (Mon)

1. Assembly
2. Miniprep
3. Transfer to liquid culture medium (repeat)