Team:Osaka/week1
From 2011.igem.org
(Difference between revisions)
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# Restriction digests of 1-9N,1-2M,1-14K,1-12M,1-1D,1-17P,1-1G,1-5A,2-16,2-18H,2-16N,1-3A,3-6H | # Restriction digests of 1-9N,1-2M,1-14K,1-12M,1-1D,1-17P,1-1G,1-5A,2-16,2-18H,2-16N,1-3A,3-6H | ||
# Gel electrophoresis | # Gel electrophoresis | ||
- | |||
# Ligation | # Ligation | ||
#* 01(K): 1-9N(upstream), 1-12M(downstream), 1-5A(vector) | #* 01(K): 1-9N(upstream), 1-12M(downstream), 1-5A(vector) | ||
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#* 05(K): 1-1D(upstream), 2-18H(downstream), 1-5A(vector) | #* 05(K): 1-1D(upstream), 2-18H(downstream), 1-5A(vector) | ||
#* 06(k): 1-9N(upstream), 2-16L(downstream), 1-5A(vector) | #* 06(k): 1-9N(upstream), 2-16L(downstream), 1-5A(vector) | ||
- | + | # Transformation of Registry parts (See Table 1). | |
- | + | ||
- | + | ||
- | # | + | |
{|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" | ||
|+Table2 | |+Table2 | ||
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|1-23L||<bbpart>BBa_K118023</bbpart>||A|| double terminator | |1-23L||<bbpart>BBa_K118023</bbpart>||A|| double terminator | ||
|} | |} | ||
+ | |||
+ | |||
+ | ==August 6 (Sat)== | ||
+ | # Transfer to LB culture medium:1-23L | ||
# Transformation of Ligation products | # Transformation of Ligation products |
Revision as of 14:19, 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
- 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
- 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 |
---|---|---|---|
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 |
Tomorrow we shall prepare the remainder of the reagents and inoculate SOB with o/n-incubated E. coli.
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.
- Colony check
- All parts successfully transformed
- Transfer to LB culture medium:1-9N,1-2M,1-14K,1-12M,1-1D,1-17P,1-1G,1-5A,2-16,2-18H,2-16N,1-3A,3-6H
August 4 (Thu)
- OD measurements throughout the day till required OD (0.3~0.7) was obtained.
- Completion of competent cells according to protocol.
- Miniprep of 1-9N,1-2M,1-14K,1-12M,1-1D,1-17P,1-1G,1-5A,2-16,2-18H,2-16N,1-3A,3-6H
August 5 (Fri)
- Restriction digests of 1-9N,1-2M,1-14K,1-12M,1-1D,1-17P,1-1G,1-5A,2-16,2-18H,2-16N,1-3A,3-6H
- Gel electrophoresis
- Ligation
- 01(K): 1-9N(upstream), 1-12M(downstream), 1-5A(vector)
- 02(K): 1-1D(upstream), 1-12M(downstream), 1-5A(vector)
- 03(K): 1-9N(upstream), 3-6H(downstream), 1-5A(vector)
- 04(K): 1-1D(upstream), 3-6H(downstream), 1-5A(vector)
- 05(K): 1-1D(upstream), 2-18H(downstream), 1-5A(vector)
- 06(k): 1-9N(upstream), 2-16L(downstream), 1-5A(vector)
- Transformation of Registry parts (See Table 1).
ID | Part Name | Resistance | Description |
---|---|---|---|
1-23L | <bbpart>BBa_K118023</bbpart> | A | double terminator |
August 6 (Sat)
- Transfer to LB culture medium:1-23L
- Transformation of Ligation products
- Preparation of LB plates (33 Amp, 14 Kan)