Calculate persistence length of a single dsDNA molecule

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Discussion Overview

The discussion revolves around calculating the persistence length of a single double-stranded DNA (dsDNA) molecule using experimental data from force vs. extension measurements. The context includes the application of picoNewton forces to a dsDNA molecule using optical tweezers, with participants exploring the discrepancies between calculated and expected values of persistence length.

Discussion Character

  • Technical explanation
  • Debate/contested
  • Mathematical reasoning

Main Points Raised

  • One participant describes their approach to calculating the persistence length, noting that their calculated value of approximately 2.9 nm is significantly lower than the expected value of around 50 nm.
  • The calculation method involves converting force data to F^{-1/2} and plotting it against extension, followed by linear fitting of the data points.
  • The participant cites an interpolation formula for the extension of a worm-like chain, suggesting that the y-intercept of the fitted line relates to the persistence length.
  • Another participant expresses confusion regarding the presence of a square root in the calculation of the intercept, questioning the formula's structure that includes terms like 1/4 - 1/4.
  • A later reply corrects the equation, indicating that it should involve \big(1-\frac{x}{L_0}\Big)^{-2} instead of \big(1-\frac{x}{L_0}\Big).
  • Another participant suggests that providing the equation and the plot might clarify the situation, indicating that further information could help resolve the issue.

Areas of Agreement / Disagreement

Participants do not appear to reach a consensus on the correct application of the formula or the interpretation of the results, with some expressing confusion and others attempting to clarify the equations involved.

Contextual Notes

There are unresolved aspects regarding the assumptions made in the calculations, the choice of force ranges, and the interpretation of the fitted data. The discussion reflects uncertainty about the correct formulation and its implications for the persistence length calculation.

pen
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Hello!

1. The problem statement and all variables

AIM: Calculating persistence length P of a single dsDNA molecule from a data set of force F (to the molecule) vs. extension x measurements. Experimental background: pN forces were applied to a single dsDNA molecule spanned between two μm-beads using an optical tweezer.

PROBLEM: When I calculate P, I get values of about 2.9 nm, which is far below the expected value for P of dsDNA, which is about 50nm.

Homework Equations



The calculation was done as follows: For a chosen force range the F-data were converted to F^{-1/2} and plotted vs. x. The data points were fitted linearly.

According to an interpolation formula the extension x of a worm like chain with contour length L_0 (Bustamante et al.,1994) is:

\frac{FP}{k_BT}=\frac{1}{4}(1−\frac{x}{L_0})−2−\frac{1}{4}+\frac{x}{L_0},

applicable for a force range of ~5-15pN, where the molecule reveals a linear F-x relationship (like a Hookean spring).

From that follows that the y-intercept of the straight line fitted to the data points is 2\sqrt{\frac{P}{k_BT}}.

The Attempt at a Solution



The problem is (as I think) that the slope of the fitted straight line is too low. So I chose different force-ranges, as I thought, that the chosen force range might be wrong. But that didnt work. In the attachement of the thread "Calculate persistence length from force extension data of a single DNA" one can find the force curve and a F^{-1/2}-x graph, plotted for a force range of 6-16pN, with the linear fit: slope of -1.5151, the y-intercept at 1.6931 and a calculated persistence length of 2.9455 nm

I really would appreciate some help

Pen
 
Last edited:
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pen said:
Hello!

1. The problem statement and all variables

AIM: Calculating persistence length P of a single dsDNA molecule from a data set of force F (to the molecule) vs. extension x measurements. Experimental background: pN forces were applied to a single dsDNA molecule spanned between two μm-beads using an optical tweezer.

PROBLEM: When I calculate P, I get values of about 2.9 nm, which is far below the expected value for P of dsDNA, which is about 50nm.

Homework Equations



The calculation was done as follows: For a chosen force range the F-data were converted to F^{-1/2} and plotted vs. x. The data points were fitted linearly.

According to an interpolation formula the extension x of a worm like chain with contour length L_0 (Bustamante et al.,1994) is:

\frac{FP}{k_BT}=\frac{1}{4}(1−\frac{x}{L_0})−2−\frac{1}{4}+\frac{x}{L_0},

applicable for a force range of ~5-15pN, where the molecule reveals a linear F-x relationship (like a Hookean spring).

From that follows that the y-intercept of the straight line fitted to the data points is 2\sqrt{\frac{P}{k_BT}}.

The Attempt at a Solution



The problem is (as I think) that the slope of the fitted straight line is too low. So I chose different force-ranges, as I thought, that the chosen force range might be wrong. But that didnt work. In the attachement of the thread "Calculate persistence length from force extension data of a single DNA" one can find the force curve and a F^{-1/2}-x graph, plotted for a force range of 6-16pN, with the linear fit: slope of -1.5151, the y-intercept at 1.6931 and a calculated persistence length of 2.9455 nm

I really would appreciate some help

Pen

I know approximately nothing about this subject, but the equation quoted relating F and x which I gather are your experimental parameters is linear so I don't understand how a square root could enter your calculation of an intercept. I wonder at a formula that contains 1/4 - 1/4.
 
Hello Epenguin!

Sorry, my mistake: it should be \big(1-\frac{x}{L_0}\Big)^{-2} instead of \big(1-\frac{x}{L_0}\Big) in the equation.
 
Too much guessing needed now, maybe if you quoted the equation and showed the plot something might become apparent.
 

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