Measuring Oligonucleotide Hydrolysis in Cellular Lysates via Viscosity Measurements.

Journal Article (Journal Article)

Cell lysis, a process that releases host oligonucleotides, is required in many biotechnological applications. However, intact oligonucleotides in crude cellular lysates increase the viscosity of lysates, which complicates downstream processes and routine laboratory workflows. To address this, nucleases that hydrolyze the intact oligonucleotides are commonly added, either as purified enzymes or co-expressed in genetically engineered bacterial strains. To measure oligonucleotide hydrolysis, common DNA quantification methods, such as qPCR or fluorescence-based, require expensive reagents and equipment, and cannot distinguish different-sized DNA fragments. Here, we outline a simple alternative method for measuring DNA/RNA hydrolysis in cellular lysates, by measuring their viscosity. This method only requires common laboratory supplies and a cell phone camera.

Full Text

Duke Authors

Cited Authors

  • Menacho-Melgar, R; Lynch, MD

Published Date

  • January 20, 2022

Published In

Volume / Issue

  • 12 / 2

Start / End Page

  • e4304 -

PubMed ID

  • 35127994

Pubmed Central ID

  • PMC8799667

Electronic International Standard Serial Number (EISSN)

  • 2331-8325

International Standard Serial Number (ISSN)

  • 2331-8325

Digital Object Identifier (DOI)

  • 10.21769/bioprotoc.4304

Language

  • eng