Difference between revisions of "Team:NYMU-Taipei/InterLab"

 
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<h2 id="1" class="subtitle">ANDROGENETIC ALOPECIA</h2>
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<h2 id="1" class="subtitle">Introduction</h2>
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<p>Androgenetic alopecia (AGA), or commonly referred to as male-pattern baldness, is a type of hair-loss in a well-defined pattern that mostly occurs in men. In men, this specific hair loss pattern leads to the formation of an “M” shaped hairline, and gradually proceeds to partial or complete baldness, while women generally suffer thinning of hair . Symptoms of AGA may start from a young age, and the risk increases with age. The prevalence of AGA in men above 50 years old is higher than 50%, dependent of countries and regions.</p>
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<p>The Interlab study is aimed toward developing a robust measurement procedure for GFP expression. The interlab participants follow a detailed protocol for transformation of plasmids, calibration, and measurement of the plasmid expression. For more detailed information please take a look at the official iGEM interlab measurement page. Below is the methods used and data collected during the Interlab Study: </p>  
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<p>The current understanding of the mechanism underlying AGA shows that AGA is highly related to dihydrotestosterone (DHT). DHT is an androgen hormone formed from the catalyzation of testosterone via 5alpha-reductase. The androgen receptors in the dermal papilla (DP) cells would bind to DHT, and thereby upregulate the expression of dickkopf 1 (DKK-1). DKK-1 would in turn be released from DP to follicular epithelial cells . The low-density-lipoprotein-related protein 5/6 (LRP5/6) and Kremen protein on follicular epithelial cells then bind to the secreted DKK-1, resulting in the inhibition of Wnt/ß-catenin pathway. The inhibition of Wnt/ß-catenin pathway inhibits the growth of root sheath cells in hair follicles . This process eventually shortens the anagen phase of hair growth and causes miniaturization of the hair follicle, leading to hair loss.</p>  
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<h3>Plate Well Pattern</h3>
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<p>This is the well plate pattern used in the Interlab for the final absorbance and fluorescence measurement.</p>
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<h3>OD600 Reference Point</h3>
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<p>A 96 well plate containing 4 samples of Ludox CL-X and 4 samples of distilled H2O are measured to establish and OD600 Reference Point which will later be used in the final absorbance measurement. </p>
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<h2 id="2" class="subtitle">Particle Standard Curves</h2>
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<p>A 96 well plate containing 4 replicate serial dilutions of silica beads are used to construct a standard curve of particle concentration which can be used to convert Abs600 measurements to an estimated number of cells. Below is the data and graphs collected for the particle standard curves:</p>
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<h2 id="3" class="subtitle">Fluorescein Standard Curve</h2>
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<p>A 96 well plate containing 4 replicate serial dilutions of Fluorescein. The data and standard curve below is used to compare the fluorescence data collected for the GFP between the teams.</p>
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<img src="https://static.igem.org/mediawiki/2018/3/33/T--NYMU-Taipei--interlab-form3-1.png">
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<img src="https://static.igem.org/mediawiki/2018/0/02/T--NYMU-Taipei--interlab-form3-2.png">
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<h2 id="4" class="subtitle">Abs600 Raw Plate Readings</h2>
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<p>A 96 well plate containing 4 replicates of each colony and device are measured at 0 and 6 Hour for absorbance and fluorescence. Below is the data for the fluorescence and absorbance of our devices at 0 and 6 hours.</p>
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<img src="https://static.igem.org/mediawiki/2018/3/30/T--NYMU-Taipei--interlab-form4-1.png">
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<h2 id="5" class="subtitle">Fluorescence Raw Plate Readings</h2>
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<img src="https://static.igem.org/mediawiki/2018/2/2e/T--NYMU-Taipei--interlab-form5-1.png">
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<img src="https://static.igem.org/mediawiki/2018/9/92/T--NYMU-Taipei--interlab-form5-2.png">
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<p>The devices and data that are mentioned in this webpage are collected and produced by Jake Yang, Annabel Chang, and Yasmin Lin.</p>
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Latest revision as of 07:11, 27 August 2018

HAIR TO STAY
TEAM
Team Members
Collaborations
PROJECT
Description
Design
Experiments & Results
Notebook
InterLab
Model
Demonstrate
Improve
Attributions
PARTS
Parts Overview
Basic Parts
Composite Parts
Part Collection
SAFETY
HUMAN PRACTICES
Human Practices
Education & Engagement
AWARDS
Applied Design
Entrepreneurship
Model
JUDGING FORM ⇗



Introduction

The Interlab study is aimed toward developing a robust measurement procedure for GFP expression. The interlab participants follow a detailed protocol for transformation of plasmids, calibration, and measurement of the plasmid expression. For more detailed information please take a look at the official iGEM interlab measurement page. Below is the methods used and data collected during the Interlab Study:

Plate Well Pattern

This is the well plate pattern used in the Interlab for the final absorbance and fluorescence measurement.

OD600 Reference Point

A 96 well plate containing 4 samples of Ludox CL-X and 4 samples of distilled H2O are measured to establish and OD600 Reference Point which will later be used in the final absorbance measurement.

Particle Standard Curves

A 96 well plate containing 4 replicate serial dilutions of silica beads are used to construct a standard curve of particle concentration which can be used to convert Abs600 measurements to an estimated number of cells. Below is the data and graphs collected for the particle standard curves:

Fluorescein Standard Curve

A 96 well plate containing 4 replicate serial dilutions of Fluorescein. The data and standard curve below is used to compare the fluorescence data collected for the GFP between the teams.

Abs600 Raw Plate Readings

A 96 well plate containing 4 replicates of each colony and device are measured at 0 and 6 Hour for absorbance and fluorescence. Below is the data for the fluorescence and absorbance of our devices at 0 and 6 hours.

Fluorescence Raw Plate Readings

The devices and data that are mentioned in this webpage are collected and produced by Jake Yang, Annabel Chang, and Yasmin Lin.