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

  • Thread starter dronee
  • Start date
  • Tags
    Curve Dna
In summary, the discussion revolves around using Sybr GreenI dye to monitor DNA melting, with a focus on the DNA melt curve for a sample of E. coli DNA mixed with Sybr GreenI. The curve is expected to have an S shape, with higher % (G+C) content resulting in a higher melting point. The topic also touches on the effect of a buffer with pH>9 on the DNA melt curve.
  • #1
dronee
2
0

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?
 
Last edited by a moderator:
Physics news on Phys.org
  • #2
bump. anyone?
 

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

1. What is a DNA melt curve?

A DNA melt curve is a graphical representation of the relationship between the temperature and the absorbance of DNA in a solution. It is used to analyze the stability and melting behavior of double-stranded DNA molecules.

2. How is a DNA melt curve performed?

To perform a DNA melt curve, a sample of double-stranded DNA is gradually heated while measuring the absorbance at a specific wavelength. The absorbance decreases as the DNA strands separate and the double helix structure melts.

3. What is the purpose of a DNA melt curve analysis?

A DNA melt curve analysis can provide information about the stability and purity of DNA samples, as well as the presence of mutations or variations in DNA sequences. It is commonly used in genetic research and diagnostic testing.

4. What factors can affect the shape of a DNA melt curve?

The shape of a DNA melt curve can be affected by factors such as the length and sequence of the DNA, the presence of impurities or contaminants, the concentration and type of buffer used, and the heating rate and temperature range used in the analysis.

5. How is a DNA melt curve used in PCR?

In PCR (polymerase chain reaction), a DNA melt curve is used to determine the specificity and efficiency of the amplification process. By analyzing the shape and melting temperature of the PCR product, researchers can confirm the presence of the desired DNA sequence and identify any potential contamination or non-specific amplification.

Similar threads

I Request for Input: 2D Minkowski Spacetime Diagram Generator
    • Special and General Relativity
Replies
6
Views
1K
Mathematica This Week's Finds in Mathematical Physics (Week 261)
    • MATLAB, Maple, Mathematica, LaTeX
Replies
5
Views
2K
Mathematica This Week's Finds in Mathematical Physics (Week 257)
    • MATLAB, Maple, Mathematica, LaTeX
Replies
5
Views
3K
Mathematica This Week's Finds in Mathematical Physics (Week 267)
    • MATLAB, Maple, Mathematica, LaTeX
Replies
1
Views
2K

Hot Threads

Recent Insights

Back
Top