Team:Osaka/Notebook week2

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Revision as of 09:49, 2 October 2010

Week2 : August 1 - August 7

August 2 (Mon)

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

Culture medium preparation
- LB agar plates (49 antibiotic-less plates)
- LB liquid medium (500 ml)
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

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:

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

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

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

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	(" ")

Meeting

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 {{Osaka/Calendar}}
 <html>
-<h2 class="margin">Week2:August 1 - August 7</h2>
+<h2 class="margin">Week2 : August 1 - August 7</h2>
 </html>
 <html><h3 id="0802">August 2 (Mon)</h3></html>

July
	Su	Mo	Tu	We	Th	Fr	Sa
					1	2	3
	4	5	6	7	8	9	10
	11	12	13	14	15	16	17
	18	19	20	21	22	23	24
Week1	25	26	27	28	29	30	31

August
	Su	Mo	Tu	We	Th	Fr	Sa
Week2	1	2	3	4	5	6	7
Week3	8	9	10	11	12	13	14
Week4	15	16	17	18	19	20	21
Week5	22	23	24	25	26	27	28
Week6	29	30	31

September
	Su	Mo	Tu	We	Th	Fr	Sa
Week6				1	2	3	4
Week7	5	6	7	8	9	10	11
Week8	12	13	14	15	16	17	18
Week9	19	20	21	22	23	24	25
Week10	26	27	28	29	30

October
	Su	Mo	Tu	We	Th	Fr	Sa
Week10						1	2
Week11	3	4	5	6	7	8	9
Week12	10	11	12	13	14	15	16
Week13	17	18	19	20	21	22	23
Week14	24	25	26	27	28	29	30
	31