Team:Osaka/week1

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Revision as of 13:44, 11 August 2011

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
- 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.

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
ID	Part Name	Resistance	Description
1-9N	<bbpart>BBa_K118023</bbpart>	A	RecA promoter
1-2M	<bbpart>BBa_K118022</bbpart>	A	RBS
1-14K	<bbpart>BBa_B0034</bbpart>	A	GFP(wild type)
1-12M	<bbpart>BBa_B0010</bbpart>	A	RBS+GFP+double terminator
1-1D	<bbpart>BBa_R0010</bbpart>	A	promoter(lacI regulation)
3-6H	<bbpart>BBa_E1010</bbpart>	A	CrtE I B with RBS
2-18H	<bbpart>BBa_E1010</bbpart>	A	RBS+CrtE
2-16N	<bbpart>BBa_E1010</bbpart>	A	RBS+CrtB
2-18H	<bbpart>BBa_E1010</bbpart>	A	RBS+CrtI
1-17P	<bbpart>BBa_E1010</bbpart>	A	RBS+CrtY
1-1G	<bbpart>BBa_E1010</bbpart>	A	construction plasmid containing mRFP coding device
1-3A	<bbpart>BBa_E1010</bbpart>	C	construction plasmid containing mRFP coding device
1-5A	<bbpart>BBa_E1010</bbpart>	K	construction plasmid containing mRFP coding device

August 3 (Wed)

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)

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).