Developing an insert to protect large DNA molecules during cell lysis

Determining large structural variations is difficult and time-consuming without long molecules to aid in the genome assembly. One issue is the fragility of large DNA molecules during routine molecular biology techniques. A novel inverted agarose insert was created to protect and make accessing large DNA easier. The inverted agarose insert has the cell solution in the middle and the agarose on the outside rather than the cells embedded in the agarose (plug inserts). Multiple agarose concentrations were tested to determine the best percentage of agarose to create the inverted insert. A proof-of-principle inverted insert was tested with <i>S. cerevisiae</i> cells to show that cells could be lysed inside the inverted insert, allowing the DNA to remain at full length in the middle.

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Publication Details

Journal
BioTechniques
Published
2025-03-04
DOI
https://doi.org/10.1080/07366205.2025.2483601
Primary Topic
DNA and Nucleic Acid Chemistry
Type
article
Field-Weighted Citation Impact
0.00
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article

Developing an insert to protect large DNA molecules during cell lysis

Kristy L. Kounovsky‐Shafer, Ernesto Ortíz, Martin Lopez, Charles Polen et al.
BioTechniques
DNA and Nucleic Acid Chemistry
article

Developing an insert to protect large DNA molecules during cell lysis

Kristy L. Kounovsky‐Shafer, Ernesto Ortíz, Martin Lopez, Charles Polen, Cordelia Harbison, Nate Garringer
article en

Abstract

Determining large structural variations is difficult and time-consuming without long molecules to aid in the genome assembly. One issue is the fragility of large DNA molecules during routine molecular biology techniques. A novel inverted agarose insert was created to protect and make accessing large DNA easier. The inverted agarose insert has the cell solution in the middle and the agarose on the outside rather than the cells embedded in the agarose (plug inserts). Multiple agarose concentrations were tested to determine the best percentage of agarose to create the inverted insert. A proof-of-principle inverted insert was tested with <i>S. cerevisiae</i> cells to show that cells could be lysed inside the inverted insert, allowing the DNA to remain at full length in the middle.

BioTechniquesVol. 77(3)
University of Nebraska–Lincoln (US)
Openalex Percentile: Top 95%
DNA and Nucleic Acid Chemistry
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