Team:Osaka/week1
From 2011.igem.org
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+ | {{Osaka}} | ||
+ | __NOTOC__ | ||
==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). | ||
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#* Preparation of glucose solution for making SOC medium. | #* 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 | ||
- | |||
# 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. | ||
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!ID!!Part Name!!Resistance!!Description | !ID!!Part Name!!Resistance!!Description | ||
|- | |- | ||
- | |1-9N||<bbpart> | + | |1-9N||<bbpart>BBa_J22106</bbpart>||A|| rec A (SOS) Promoter |
|- | |- | ||
|1-2M||<bbpart>BBa_K118022</bbpart>||A||RBS | |1-2M||<bbpart>BBa_K118022</bbpart>||A||RBS | ||
|- | |- | ||
- | |1-14K||<bbpart> | + | |1-14K||<bbpart>BBa_B0040</bbpart>||A||green fluorescent protein derived from jellyfish Aequeora victoria wild-type GFP |
|- | |- | ||
- | |1-12M||<bbpart> | + | |1-12M||<bbpart>BBa_E0240</bbpart>||A||GFP generator |
|- | |- | ||
- | |1-1D||<bbpart>BBa_R0010</bbpart>||A||promoter(lacI | + | |1-1D||<bbpart>BBa_R0010</bbpart>||A||promoter(lacI regulatd) |
|- | |- | ||
- | |3-6H||<bbpart> | + | |3-6H||<bbpart>BBa_K274100</bbpart>||A||CrtEBI with rbs |
|- | |- | ||
- | |2-18H||<bbpart> | + | |2-18H||<bbpart>BBa_K118014</bbpart>||A||rbs+crtE (geranylgeranyl pyrophosphate synthase) |
|- | |- | ||
- | |2-16N||<bbpart> | + | |2-16N||<bbpart>BBa_K118006</bbpart>||A||rbs+crtB (phytoene synthase) |
|- | |- | ||
- | |2-18H||<bbpart> | + | |2-18H||<bbpart>BBa_K118005</bbpart>||A||rbs+crtI (phytoene dehydrogenase) |
|- | |- | ||
- | |1-17P||<bbpart> | + | |1-17P||<bbpart>BBa_I742155</bbpart>||A||crtY (lycopene cyclase) with native rbs |
|- | |- | ||
- | |1-1G||<bbpart> | + | |1-1G||<bbpart>BBa_J04450</bbpart>||A||construction plasmid containing mRFP coding device |
|- | |- | ||
- | |1-3A||<bbpart> | + | |1-3A||<bbpart>BBa_J04450</bbpart>||C||construction plasmid containing mRFP coding device |
|- | |- | ||
- | |1-5A||<bbpart> | + | |1-5A||<bbpart>BBa_J04450</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. | Tomorrow we shall prepare the remainder of the reagents and inoculate SOB with o/n-incubated E. coli. | ||
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#* 04(K): 1-1D(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) | #* 05(K): 1-1D(upstream), 2-18H(downstream), 1-5A(vector) | ||
- | #* 06( | + | #* 06(K): 1-9N(upstream), 2-16L(downstream), 1-5A(vector) |
- | # Transformation of Registry parts (See Table | + | # Transformation of Registry parts (See Table 2). |
{|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 | ||
!ID!!Part Name!!Resistance!!Description | !ID!!Part Name!!Resistance!!Description | ||
|- | |- | ||
- | |1-23L||<bbpart> | + | |1-23L||<bbpart>BBa_B0015</bbpart>||A|| double terminator |
|} | |} | ||
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==August 6 (Sat)== | ==August 6 (Sat)== | ||
# Transfer to LB culture medium:1-23L | # Transfer to LB culture medium:1-23L | ||
- | |||
# Transformation of Ligation products | # Transformation of Ligation products | ||
# Preparation of LB plates (33 Amp, 14 Kan) | # Preparation of LB plates (33 Amp, 14 Kan) | ||
+ | |||
[[Team:Osaka/Notebook|Back to Notebook]] | [[Team:Osaka/Notebook|Back to Notebook]] | ||
+ | |||
+ | week '''1'''|[[Team:Osaka/week2|2]]|[[Team:Osaka/week3|3]]|[[Team:Osaka/week4|4]]|[[Team:Osaka/week5|5]]|[[Team:Osaka/week6|6]]|[[Team:Osaka/week7|7]]|[[Team:Osaka/week8|8]]|[[Team:Osaka/week9|9]]|[[Team:Osaka/week10|10]]|[[Team:Osaka/week11|11]]|[[Team:Osaka/week12|12]]|[[Team:Osaka/week13|13]] |
Latest revision as of 05:53, 19 October 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
- 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_J22106</bbpart> | A | rec A (SOS) Promoter |
1-2M | <bbpart>BBa_K118022</bbpart> | A | RBS |
1-14K | <bbpart>BBa_B0040</bbpart> | A | green fluorescent protein derived from jellyfish Aequeora victoria wild-type GFP |
1-12M | <bbpart>BBa_E0240</bbpart> | A | GFP generator |
1-1D | <bbpart>BBa_R0010</bbpart> | A | promoter(lacI regulatd) |
3-6H | <bbpart>BBa_K274100</bbpart> | A | CrtEBI with rbs |
2-18H | <bbpart>BBa_K118014</bbpart> | A | rbs+crtE (geranylgeranyl pyrophosphate synthase) |
2-16N | <bbpart>BBa_K118006</bbpart> | A | rbs+crtB (phytoene synthase) |
2-18H | <bbpart>BBa_K118005</bbpart> | A | rbs+crtI (phytoene dehydrogenase) |
1-17P | <bbpart>BBa_I742155</bbpart> | A | crtY (lycopene cyclase) with native rbs |
1-1G | <bbpart>BBa_J04450</bbpart> | A | construction plasmid containing mRFP coding device |
1-3A | <bbpart>BBa_J04450</bbpart> | C | construction plasmid containing mRFP coding device |
1-5A | <bbpart>BBa_J04450</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 2).
ID | Part Name | Resistance | Description |
---|---|---|---|
1-23L | <bbpart>BBa_B0015</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)
week 1|2|3|4|5|6|7|8|9|10|11|12|13