Tokyo/IPTG assay

From 2007.igem.org

(Difference between revisions)
Line 1: Line 1:
__NOTOC__
__NOTOC__
 +
 +
==[[Tokyo_Tech|Abstract]]  [[Tokyo/Model|Concept & Model]]  [[Tokyo/Requirements |Requirements]]  [[Tokyo/Genetic circuit|Genetic_circuit]]  [[Tokyo/Works|Works]]  [[Tokyo/About our team|About_our_team]]==
 +
<br>[[Tokyo/Works|Works top]]  0.[[Tokyo/Works/Hybrid promoter|Hybrid promoter]]  1.[[Tokyo/Works/Formulation |Formulation]]  2.[[Tokyo/Works/Assay |Assay1]]  3.[[Tokyo/Works/Simulation |Simulation]]  4.[[Tokyo/Works/Assay2 |Assay2]]  5.[[Tokyo/Works/Future works |Future works]]
 +
== IPTG assay ==
== IPTG assay ==
===Purpose: ===
===Purpose: ===

Revision as of 21:43, 24 October 2007


Abstract  Concept & Model  Requirements  Genetic_circuit  Works  About_our_team


Works top  0.Hybrid promoter  1.Formulation  2.Assay1  3.Simulation  4.Assay2  5.Future works

IPTG assay

Purpose:


To determine the order of the concentration of IPTG necessary for the activation of our lux-lac hybrid promoter in the LacI producing pTrc99A cells.
The order of the concentration is used for more detailed assay with narrower range of the IPTG concentration.

Samples:


A4 placQI in pTrc99A
A4 ΔP in pTrc99A
Lux-lac hybrid promter + A4 in pBR322
Lux-lac hybrid promter + A4 in pTrc99A

Procedure:

Fig.1: Exogenously added IPTG


prepare overnight culture for each sample
make fresh culture
take 3 ul of the overinight culture into 3 ml of LB (Amp and/or Kan) in Falcon tubes.
incubate for 2 to 3 hours until the observed OD is around 0.5
add AHL & IPTG solution
[AHL]final (in 3 ml LB culture) = 10 nM
[IPTG]final (in 3 ml LB culture) = 1000, 100, 10, 1, 0.1, 0.01, and 0 mM
incubate for 2 to 3 hours
apply 150 ul of samples into 96-well plaste
FLA measurement

Result & Conclusion:

Fig.2: IPTG assay graph