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