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Team:NTU-Singapore/Notebook
From 2009.igem.org
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* PE: pNO-GFP; J23101-GFP; J23119-GFP; HO1 | * PE: pNO-GFP; J23101-GFP; J23119-GFP; HO1 | ||
* DD: HO1 | * DD: HO1 | ||
| + | |||
| + | |||
| + | <u>'''7th July'''</u> | ||
| + | * PE: CHE, IFP, placIQ | ||
| + | * DD:CHE-T, IFP-T. HO1, Term | ||
| + | * Ligation | ||
| + | |||
| + | |||
| + | <u>'''8th July'''</u> | ||
| + | * DD: pNO-GFP; J23101-GFP; J23119-GFP | ||
| + | |||
| + | |||
| + | <u>'''9th July'''</u> | ||
| + | * PE | ||
| + | * DD: ptet. r-CHE-T, r-HO1, r-IFP-T | ||
| + | * Ligation: r-HO1 +r-IFP-T | ||
| + | |||
| + | |||
| + | <u>'''10th July'''</u> | ||
| + | * PE | ||
| + | * DD: ptet; r-CHE-T, Term, tetR | ||
| + | |||
| + | |||
| + | <u>'''11th July'''</u> | ||
| + | * PE | ||
| + | * DD: r-CHE-T; r-HO1-r-IFP-T;CHE-T; r-CHE-T | ||
| + | |||
| + | |||
| + | <u>'''13th July'''</u> | ||
| + | * DD: ptet; r-CHE-T; r-HO1-r-IFP-T; TetR; Term | ||
| + | * Ligation: ptet + r-CHE-T; ptet + r-HO1-r-IFP-T; TetR + Term | ||
| + | |||
| + | |||
| + | <u>'''15th July'''</u> | ||
| + | * PE | ||
| + | * DD: pTet; rbs; IFP; CHE; TetR-T | ||
| + | |||
| + | |||
| + | <u>'''16th July'''</u> | ||
| + | * DD: pTet; r-CHE-T; Inv; r-HO1-r-IFP-T; Term; TetR | ||
| + | * Ligation | ||
| + | |||
| + | |||
| + | <u>'''18th July'''</u> | ||
| + | * PE | ||
| + | * DD and Single Digestion (SD) :r-HO1-r-IFP-T (because they have the same size) | ||
| + | |||
| + | |||
| + | <u>'''20th July'''</u> | ||
| + | * DD:pTet; J23119; r-HO1-r-IFP-T; TetR; Term; placIQ; inv | ||
| + | * Ligation: pTet +r-HO1-r-IFP-T; J23119 + r-HO1-r-IFP-T; placIQ +Inv; TetR +Term | ||
| + | * PE | ||
| + | |||
| + | |||
| + | <u>'''23rd July'''</u> | ||
| + | * PE | ||
| + | * DD:J23119; Inv; r-HO1; r-GFP-T; pTet; placIQ-Inv; TetR-T | ||
| + | |||
| + | |||
| + | <u>'''24th July'''</u> | ||
| + | * PE | ||
| + | * DD: J23119; r-CHE-T; pTet; inv;r-HO1-r-IFP-T; ptet-IFP | ||
| + | * Ligation: J23119+r-CHE-T; ptet+r-CHE-T; ptet+r-HO1-r-IFP-T; J23119+Inv | ||
| + | |||
| + | |||
| + | <u>'''28th July'''</u> | ||
| + | * PE | ||
| + | * DD: r-HO1; r-GFP-T; J23119;r-CHE-T;r-HO1-r-IFP-t; ptet; r-CHE-T | ||
| + | * Ligation: r-HO1 +r-GFP-T; J23119 +r-CHE-T; J23119 + r-HO1-r-IFP-T; pTet +r-CHE-T; pTet +r-GFP-T | ||
| + | |||
| + | |||
| + | <u>'''30th July'''</u> | ||
| + | * PE | ||
| + | *DD : J23119-r-CHE-T; ptet-r-CHE-T; J23119-r-HO1-r-IFP-T; ptet-r-HO1-r-IFP-T; r-HO1-r-GFP-T; J23119; pTet; TetR; rbs | ||
| + | * Ligation: r-HO1 +r-GFP-T; J23119 +r-CHE-T; J23119+r-HO1-IFP-T; ptet+r-CHE-T; ptet+r-GFP-T | ||
| + | |||
| + | |||
| + | == August == | ||
| + | |||
| + | |||
| + | <u>''' '''</u> | ||
| + | * | ||
| + | |||
| + | |||
| + | <u>''' '''</u> | ||
| + | * | ||
| + | |||
| + | <u>''' '''</u> | ||
| + | * | ||
| + | |||
<u>''' '''</u> | <u>''' '''</u> | ||
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<u>''' '''</u> | <u>''' '''</u> | ||
* | * | ||
| + | |||
| + | |||
| + | <u>''' '''</u> | ||
| + | * | ||
| + | |||
| + | |||
| + | |||
| + | |||
Revision as of 19:21, 21 October 2009
Experiment Timeline
June
18th June
- Preparation of LB medium
- Amplification of parts from registry (pTet, TetR, HO1, Low copy plasmid)
19th June
- Antibiotic stock preparation
- Inoculation
20th June
- Our first Plasmid Extraction (PE)!
22nd June
- First Double Digestion (DD) and Gel Check!
23rd June
- PE & DD of the registry constructs….
24th June
- Our direct synthesized parts finally arrived. We now have CHE (cholesterol esterase), IFP(Infrared protein 1.4) and pNO (NO sensing promoter).
26th June
- PE , DD and Gel check for the direct synthesized gene sequence.
27th June
- Gel Check for CHE, IFP and pNO
July
1st July
- PE for plaqIQ, J23101, J23119, B0034(rbs), B0015(Term-T for short), B0034-GFP-B0015
2nd July
- PE for pNO, IFP, CHE
- DD of pNO, J23119, J23101, rbs-GFP-T
- Our first ligation!
3rd July
- DD of CHE, IFP and B0015, plaqIQ and plux.
- Ligation.
4th July
- PE: pNO-GFP; J23101-GFP; J23119-GFP; HO1
- DD: HO1
7th July
- PE: CHE, IFP, placIQ
- DD:CHE-T, IFP-T. HO1, Term
- Ligation
8th July
- DD: pNO-GFP; J23101-GFP; J23119-GFP
9th July
- PE
- DD: ptet. r-CHE-T, r-HO1, r-IFP-T
- Ligation: r-HO1 +r-IFP-T
10th July
- PE
- DD: ptet; r-CHE-T, Term, tetR
11th July
- PE
- DD: r-CHE-T; r-HO1-r-IFP-T;CHE-T; r-CHE-T
13th July
- DD: ptet; r-CHE-T; r-HO1-r-IFP-T; TetR; Term
- Ligation: ptet + r-CHE-T; ptet + r-HO1-r-IFP-T; TetR + Term
15th July
- PE
- DD: pTet; rbs; IFP; CHE; TetR-T
16th July
- DD: pTet; r-CHE-T; Inv; r-HO1-r-IFP-T; Term; TetR
- Ligation
18th July
- PE
- DD and Single Digestion (SD) :r-HO1-r-IFP-T (because they have the same size)
20th July
- DD:pTet; J23119; r-HO1-r-IFP-T; TetR; Term; placIQ; inv
- Ligation: pTet +r-HO1-r-IFP-T; J23119 + r-HO1-r-IFP-T; placIQ +Inv; TetR +Term
- PE
23rd July
- PE
- DD:J23119; Inv; r-HO1; r-GFP-T; pTet; placIQ-Inv; TetR-T
24th July
- PE
- DD: J23119; r-CHE-T; pTet; inv;r-HO1-r-IFP-T; ptet-IFP
- Ligation: J23119+r-CHE-T; ptet+r-CHE-T; ptet+r-HO1-r-IFP-T; J23119+Inv
28th July
- PE
- DD: r-HO1; r-GFP-T; J23119;r-CHE-T;r-HO1-r-IFP-t; ptet; r-CHE-T
- Ligation: r-HO1 +r-GFP-T; J23119 +r-CHE-T; J23119 + r-HO1-r-IFP-T; pTet +r-CHE-T; pTet +r-GFP-T
30th July
- PE
- DD : J23119-r-CHE-T; ptet-r-CHE-T; J23119-r-HO1-r-IFP-T; ptet-r-HO1-r-IFP-T; r-HO1-r-GFP-T; J23119; pTet; TetR; rbs
- Ligation: r-HO1 +r-GFP-T; J23119 +r-CHE-T; J23119+r-HO1-IFP-T; ptet+r-CHE-T; ptet+r-GFP-T
August
