DNA Melting Curves: E. coli and Low % GC Content Samples with Sybr GreenI Dye

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The discussion focuses on creating DNA melting curves using Sybr GreenI dye to analyze E. coli DNA and samples with lower GC content. Participants are asked to plot the melting curve, which should resemble an S-shape, with temperature on the x-axis and fluorescence on the y-axis. It is noted that samples with higher GC content will have higher melting points, shifting the curve upwards. There is uncertainty regarding the melting curve for DNA dissolved in a buffer at pH > 9, indicating a need for further clarification. Overall, the conversation emphasizes understanding the relationship between GC content and DNA melting behavior.
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Homework Statement



You have a dye, Sybr GreenI which binds only to double stranded DNA (not single stranded). Once bound, it fluoresces strongly and can be used to monitor DNA melting (transition from double stranded to single stranded).

(i) on the axes provided draw the DNA melt curve for a sample of E. coli DNA which has been mixed with Sybr GreenI.
--> the axes shown has Temperature for x-axis and Fluorescence for y-axis.
on the same graph show the melt curve for a sample of DNA
(ii) with a lower %(G+C) content
(iii) from the same source dissolved in a buffer at pH>9 prior to the measurements.

Homework Equations





The Attempt at a Solution



(i) okay so I am thinking the shape of the curve is like an S
kind of like this: http://www.entelechon.com/wp-content/uploads/2008/10/dna_melting-300x178.gif
but is it exactly like that? or is the other way?

(ii) for this one i know that i just have to draw the exact same shape except more towards 0 since more GC content means higher melting point.

(iii) I am not sure about this one.
help please?
 
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