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Measuring Oligonucleotide Hydrolysis in Cellular Lysates via Viscosity Measurements.

Publication ,  Journal Article
Menacho-Melgar, R; Lynch, MD
Published in: Bio-protocol
January 2022

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.

Duke Scholars

Published In

Bio-protocol

DOI

EISSN

2331-8325

ISSN

2331-8325

Publication Date

January 2022

Volume

12

Issue

2

Start / End Page

e4304

Related Subject Headings

  • 32 Biomedical and clinical sciences
  • 31 Biological sciences
 

Citation

APA
Chicago
ICMJE
MLA
NLM
Menacho-Melgar, R., & Lynch, M. D. (2022). Measuring Oligonucleotide Hydrolysis in Cellular Lysates via Viscosity Measurements. Bio-Protocol, 12(2), e4304. https://doi.org/10.21769/bioprotoc.4304
Menacho-Melgar, Romel, and Michael D. Lynch. “Measuring Oligonucleotide Hydrolysis in Cellular Lysates via Viscosity Measurements.Bio-Protocol 12, no. 2 (January 2022): e4304. https://doi.org/10.21769/bioprotoc.4304.
Menacho-Melgar R, Lynch MD. Measuring Oligonucleotide Hydrolysis in Cellular Lysates via Viscosity Measurements. Bio-protocol. 2022 Jan;12(2):e4304.
Menacho-Melgar, Romel, and Michael D. Lynch. “Measuring Oligonucleotide Hydrolysis in Cellular Lysates via Viscosity Measurements.Bio-Protocol, vol. 12, no. 2, Jan. 2022, p. e4304. Epmc, doi:10.21769/bioprotoc.4304.
Menacho-Melgar R, Lynch MD. Measuring Oligonucleotide Hydrolysis in Cellular Lysates via Viscosity Measurements. Bio-protocol. 2022 Jan;12(2):e4304.

Published In

Bio-protocol

DOI

EISSN

2331-8325

ISSN

2331-8325

Publication Date

January 2022

Volume

12

Issue

2

Start / End Page

e4304

Related Subject Headings

  • 32 Biomedical and clinical sciences
  • 31 Biological sciences