iGEM 2009 — Team NTU-Singapore

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# Team:NTU-Singapore/Notebook

### From 2009.igem.org

Welcome to the notebook page!

A record of our exploits.

[Home](/Team%3ANTU-Singapore "Team:NTU-Singapore")

[pLaq*U*e Out!](/Team%3ANTU-Singapore/Project "Team:NTU-Singapore/Project")

[DIYbio](/Team%3ANTU-Singapore/HPA "Team:NTU-Singapore/HPA")

[Accomplishments!](/Team%3ANTU-Singapore/Notebook/Deliverables "Team:NTU-Singapore/Notebook/Deliverables")

**Notebook**

[About Us](/Team%3ANTU-Singapore/About/Team "Team:NTU-Singapore/About/Team")

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

**4th August**

* DD :rbs; NorR

**5th August**

* PE
* DD
* Ligation

**12th August**

* DD
* Ligation

**14th August**

* DD
* Ligation

**17th August**

* DD
* Our first SDS-PAGE—J23119-r-CHE-T; pTet-r-CHE-T

**18th August**

* DD
* Ligation

**21st August**

* DD
* Characterization of J23119

**24th August**

* PCR verification –HO1, pTet constructs

**25th August**

* pTet characterization

**26th August**

* DD

**27th August**

* DD
* Ligation

**28th August**

* PCR verification – CHE constructs
* DD
* Ligation

**29th August**

* DD
* Ligation
* PCR verification –IFP constructs

**31st August**

* PCR verification –HO1, CHE constructs
* DD
* Ligation

## September

**1st September**

* PCR verification –IFP, NorR, pNO
* DD
* Ligation

**2nd September**

* DD
* Ligation

**3rd September**

* DD
* Ligation

**4th September**

* DD
* Ligation

**6th September**

* PCR --J23119-r-NorR-T

**7th September**

* DD,
* Ligation

**8th September**

* PCR verification –IFP, HO1 constructs

**9th September**

* PCR verification –HO1 constructs

**10th September**

* DD
* Ligation

**11th September**

* PCR verification –NorR, IFP, HO1 constructs
* Ligation

**12thS eptember**

* PCR verification – NorR, IFP, HO1 constructs

**14thSeptember**

* PCR verification – IFP, HO1 constructs

**15th September**

* DD
* Ligation
* SDS-Page

**16th September**

* pNO characterization

**17th September**

* Ligation

**18th September**

* DD
* Ligation

**19th September**

* DD

**20th September**

* Ligation

**22nd September**

* PCR verification –IFP, HO1
* pNO characterization

**23rd September**

* PCR verification --CHE

**24th September**

* PCR verification –CHE, HO1
* DD
* Ligation
* Optimization of CHE characterization experiment

**26th September**

* DD
* Ligation

Navigation

**What we did**
 [Accomplishments](/Team%3ANTU-Singapore/Notebook/Deliverables "Team:NTU-Singapore/Notebook/Deliverables")
 [Safety](/Team%3ANTU-Singapore/Notebook/Safety "Team:NTU-Singapore/Notebook/Safety")
 [Our protocols](/Team%3ANTU-Singapore/Notebook/Protocols "Team:NTU-Singapore/Notebook/Protocols")
 [Brainstorming ideas](/Team%3ANTU-Singapore/Notebook/Brainstorming "Team:NTU-Singapore/Notebook/Brainstorming")
 [Online resources](/Team%3ANTU-Singapore/Notebook/Links "Team:NTU-Singapore/Notebook/Links")

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