Data from: Diverse, distinct, and densely packed DNA nanostar droplets
Data files
Feb 03, 2026 version files 5.61 GB
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2025-01-29-DiverseNanostars-DataArchive.zip
5.61 GB
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README.md
4.43 KB
Abstract
The liquid-liquid phase separation of biomolecules is an important process for intracellular organization. Biomolecular sequence combinatorics leads to a large variety of proteins and nucleic acids which can interact to form a diversity of dense liquid ('condensate') phases. The relationship between sequence design and the diversity of the resultant phases is therefore of interest. Here, we explore this question using the DNA nanostar system, which permits creation of multi-phase condensate droplets through sequence engineering of the sticky end bonds that drive particle-particle attraction. We explore the theoretical limits of nanostar phase diversity, then experimentally demonstrate the ability to create 9 distinct, non-adhering nanostar phases that do not share components. We further study how thermal processing affects the morphology and dynamics of such a highly diverse condensate system. We particularly show that a rapid temperature quench leads to the formation of a densely packed 2-D layer of droplets that is transiently stabilized by caging effects enabled by the phase diversity, leading to glassy dynamics, such as slow coarsening and dynamic heterogeneity. Generally, our work provides experimental insight into the thermodynamics of phase separation of complex mixtures, and demonstrates the rational engineering of complex, long-range, multi-phase droplet structures.
Dataset DOI: 10.5061/dryad.7d7wm387m
Description of the data and file structure
This document describes the data repository associated with the paper Diverse, Distinct, and Densely Packed DNA Nanostar Droplets by Aria S. Chaderjian, Sam Wilken, and Omar A. Saleh.
Files and variables
This repository consists of a .zip archive containing 51 data files, as well as a Mathematica notebook and its corresponding PDF.
The .zip ( 2025-01-29-DiverseNanostars-DataArchive.zip) contains the following files:
-This ReadMe file
-Three files containing all 4-, 6-, and 8-base DNA palindromes with Delta G values calculated from the 2004 SantaLucia parameters (see the Mathematica notebook GraphStructureOfOrthogonalPalindromes-ForRelease.nb). These files underlie Fig. 1D of the main text.
Filenames:
- AllFourBasePalindromesWithDG.csv
- AllSixBasePalindromesWithDG.csv
- AllEightBasePalindromesWithDG.csv
-Three files containing all cliques of the maximum size of 3-base orthogonal sticky ends with Delta G within 0.4, 0.8, and 1.2 kcal/mol of each other. The data is a list of lists of lists, namely a list containing all cliques, with each clique containing lists of sticky end sequence and corresponding Delta G value. Cliques contain 8, 9, and 10 sticky ends for deltaG spread of 0.4, 0.8, and 1.2 kcal/mol respectively. These files were generated from the Mathematica notebook GraphStructureOfOrthogonalPalindromes-ForRelease.nb.
Filenames:
- Cliques04
- Cliques08
- Cliques12
-One file 'DimerConcentrations.csv' containing the concentration of each complex that resulted from inputting all sticky end sequences with the unpaired T as listed in Fig. 1E of the main text, referred to as strands 1-9. Each sticky end was input at 12 uM each. This file underlies Fig. 1F of the main text.
-One file 'All9.ome.tif' which corresponds to Fig. 3B-D of the main text. 1 pixel = 0.4887 um. 200 ms exposure time was used with 100% LED power. The images are in the order Cy3, Cy5, Atto488. No background corrections have been performed (though they were for the analysis).
-Nine folders 'SingleDark_x.tif' which correspond to the single-dark confocal experiments where one NS phase was left unlabeled (only visible in brightfield). x in the naming scheme corresponds to the NS left unlabeled. Each folder contains 4 files, c0.tif, c1.tif, c2.tif, c3.tif, which correspond to Cy3, Atto488, Cy5, and brightfield respectively. 1 pixel = 0.22 um. 800 ms exposure time was used for Cy3 and Cy5 and 4 s exposure time for Atto488, all with 100% laser power. No background corrections have been performed (though they were for the analysis).
-One video 'PostRamp.ome.tif' which corresponds to Fig. 4A of the main text. 1 pixel = 0.733 um. 200 ms exposure time was used with 10% LED power. The video begins 40 minutes after the system reached 25C with 10 minute intervals. The images are in order (t1, c1), (t1, c2), (t1, c3), (t2, c1), (t2, c2), ... where t1 corresponds to the first timepoint, c1 corresponds to the Cy3 image, c2 corresponds to the Cy5 image, and c3 corresponds to the Atto488 image. No background corrections have been performed (though they were for the analysis).
-Six videos titled 'PostQuench_x.ome.tif' where x corresponds to the order in which the video was acquired. All 6 videos are of the same region and correspond to Fig. 4B and Fig. 5 of the main text. This was the same sample as used in PostRamp.ome.tif, and the same imaging conditions apply except that an additional 1.5x magnification setting was turned on, so 1 pixel = 0.4887 um. No background corrections have been performed. Start times (time after reaching 25C) and intervals between frames in the videos are as follows, all listed in minutes:
x=1 1 5
x=2 260 10
x=3 1190 10
x=4 2530 10
x=5 3960 20
x=6 7450 10
Code/software
A Mathematica notebook titled 'GraphStructureOfOrthogonalPalindromes-ForRelease.nb' is included, saved also as a PDF. This notebook accomplishes the graph-theoretic analysis of the set of all 4-, 6-, and 8-base palindromes, outputting lists of the largest groups of mutually orthogonal palindromes that accomplish a given spread in binding energy, Delta G. More details are included in the comments of the notebook.
