Revision as of 03:22, 18 October 2018

Results

Introduction>

To better characterize the standard curve for the EtBr-DNA Dot Blot, we are interested first whether the amount of EtBr significantly affects the fluorescent when the DNA concentration is varied between different amounts of EtBr and demonstrate there is significant difference in fluorescent between. We almost must characterize the predictive ability of the standard curve using some known DNA concentration that is not used for the curve.

Methods>

Statistical analysis was done with R-studio and the Standard Curve was generated using Excel. Data was entered into Excel and standardized using the following fomula: StandardPixel = (Value-MIN)\div(MAX-MIN)

Results>

We examined the correlation of 125ng DNA and 16ng DNA under varied conditions of EtBr. We found that there is a low correlation when between the amount of EtBr and fluorescent.

125 ng of DNA vs EtBr

Figure 1. Scatter plot of 125ng DNA with varied amount of EtBr. r=0.23<>

16 ng of DNA vs EtBr

Figure 1. Scatter plot of 16ng DNA with varied amount of EtBr. r=0.20<>

When number of DNA was varied, we found that 125ng of DNA (β = 43.0, p < .001), and 16 ng of DNA (β=41.9, p < .001) were significant predictors of pixel intensity. Zero ng of DNA was used as the reference variable. The overall model fit was R^2 = 0.57, F-statistics = 41.02, p<.001.

This goes in contrast to our Mass-Action model which shows that the amount of EtBr has a significant effect on the pixel intensity. However, it is obvious that EtBr is needed for DNA to fluorescent therefore a future study should examine at what concentration of EtBr do we see no fluorescent when the amount of DNA is varied.

R-Output for 125 ng, 16 ng, and 0 ng DNA vs EtBr

Figure 4. Standard Curve made from known concentration of DNA and analyzed using ImageJ®.>

Standardized Pixel Intensity vs DNA Concentration

Prediction of the Curve

As of now, we do not have enough samples but as more teams send their samples in for us to analyze we will report the prediction of the curve.>

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 <h3 class="default-padding" style:"color: black"><u> 125 ng of DNA vs EtBr</u></h3>
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      <center><a href="https://static.igem.org/mediawiki/2018/a/a8/T--CUNY_Kingsborough--iGEM125ngDNAvsEtBr.jpgg" target="_blank"><img src="https://static.igem.org/mediawiki/2018/a/a8/T--CUNY_Kingsborough--iGEM125ngDNAvsEtBr.jpg"></a></center>
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 <h3 class="default-padding" style:"color: black"><u> 16 ng of DNA vs EtBr</u></h3>
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      <center><a href="https://static.igem.org/mediawiki/2018/c/c7/T--CUNY_Kingsborough--iGEM16ngDNAvsEtBr.jpg" target="_blank"><img src="https://static.igem.org/mediawiki/2018/c/c7/T--CUNY_Kingsborough--iGEM16ngDNAvsEtBr.jpg"></a></center>
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 <h3 class="default-padding" style:"color: black">R-Output for 125 ng, 16 ng, and 0 ng DNA vs EtBr<u></u></h3>
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      <center><a href="https://static.igem.org/mediawiki/2018/4/4e/T--CUNY_Kingsborough--iGEMResultsStats2018.jpg" target="_blank"><img src="https://static.igem.org/mediawiki/2018/4/4e/T--CUNY_Kingsborough--iGEMResultsStats2018.jpg"></a></center>
 <p class="no-rise-padding"><b>Figure 4.</b> Standard Curve made from known concentration of DNA and analyzed using ImageJ®.></p>
 <h3 class="default-padding" style:"color: black"><u> Standardized Pixel Intensity vs DNA Concentration</u></h3>
-<div id="image-center"
      <center><a href="https://static.igem.org/mediawiki/2018/c/c7/T--CUNY_Kingsborough--iGEM16ngDNAvsEtBr.jpg" target="_blank"><img src="https://static.igem.org/mediawiki/2018/c/c7/T--CUNY_Kingsborough--iGEM16ngDNAvsEtBr.jpg"></a></center>

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