Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Concept Videos

RNA-seq03:21

RNA-seq

11.6K
RNA sequencing, or RNA-Seq, is a high-throughput sequencing technology used to study the transcriptome of a cell. Transcriptomics helps to interpret the functional elements of a genome and identify the molecular constituents of an organism. Additionally, it also helps in understanding the development of an organism and the occurrence of diseases. 
Before the discovery of RNA-seq, microarray-based methods and Sanger sequencing were used for transcriptome analysis. However, while...
11.6K
Real Time RT-PCR02:57

Real Time RT-PCR

64.3K
Real-time reverse transcription-polymerase chain reaction, or Real-time RT-PCR, is an analytical tool used to determine the expression level of target genes. The method involves converting mRNA to complementary DNA with the help of an enzyme known as reverse transcriptase, followed by the PCR amplification of the cDNA. These two processes can be performed simultaneously in a single tube or separately as a two-step reaction.
The real-time quantification of the number of amplified products is...
64.3K
Maxam-Gilbert Sequencing01:05

Maxam-Gilbert Sequencing

12.5K
In the same year as the discovery of the Sanger sequencing method, another group of scientists, Allan Maxam and Walter Gilbert, demonstrated their chemical-cleavage method for DNA sequencing. The Maxam-Gilbert method relies on using different chemicals that can cleave the DNA sequence at specific sites, the separation of resulting DNA fragments of variable size using electrophoresis, and deciphering the DNA sequence from the resulting gel bands.
Challenges of the Maxam-Gilbert Method
The...
12.5K

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

The ACS Division of Analytical Chemistry Awards Program: Honoring Our Recent Awardees.

Analytical chemistry·2025
Same author

Comparison of N-Glycopeptide to Released N-Glycan Abundances and the Influence of Glycopeptide Mass and Charge States on N-Linked Glycosylation of IgG Antibodies.

Journal of proteome research·2024
Same author

Quantification of mRNA in Lipid Nanoparticles Using Mass Spectrometry.

Analytical chemistry·2024
Same author

Quantification of Total Vitamin-D-Binding Protein and the Glycosylated Isoforms by Liquid Chromatography-Isotope Dilution Mass Spectrometry.

Journal of proteome research·2017
Same author

Development of an Improved Standard Reference Material for Vitamin D Metabolites in Human Serum.

Analytical chemistry·2017

Related Experiment Video

Updated: Jan 5, 2026

DNAzyme-dependent Analysis of rRNA 2’-O-Methylation
09:12

DNAzyme-dependent Analysis of rRNA 2’-O-Methylation

Published on: September 16, 2019

8.7K

Absolute Quantification of RNA or DNA Using Acid Hydrolysis and Mass Spectrometry.

Mark S Lowenthal1, Eva Quittman1, Karen W Phinney1

  • 1Biomolecular Measurement Division , National Institute of Standards and Technology , 100 Bureau Drive, Stop 8314 , Gaithersburg , Maryland 20899 , United States.

Analytical Chemistry
|October 23, 2019
PubMed
Summary

Accurate quantification of RNA and DNA oligomers is now possible using acid hydrolysis and isotope dilution mass spectrometry (ID-MS). This method precisely measures nucleobases, enabling reliable absolute quantification and traceability.

More Related Videos

Simultaneous Mapping and Quantitation of Ribonucleotides in Human Mitochondrial DNA
12:35

Simultaneous Mapping and Quantitation of Ribonucleotides in Human Mitochondrial DNA

Published on: November 14, 2017

9.8K
Identification of RNA Fragments Resulting from Enzymatic Degradation using MALDI-TOF Mass Spectrometry
09:20

Identification of RNA Fragments Resulting from Enzymatic Degradation using MALDI-TOF Mass Spectrometry

Published on: April 11, 2022

3.8K

Related Experiment Videos

Last Updated: Jan 5, 2026

DNAzyme-dependent Analysis of rRNA 2’-O-Methylation
09:12

DNAzyme-dependent Analysis of rRNA 2’-O-Methylation

Published on: September 16, 2019

8.7K
Simultaneous Mapping and Quantitation of Ribonucleotides in Human Mitochondrial DNA
12:35

Simultaneous Mapping and Quantitation of Ribonucleotides in Human Mitochondrial DNA

Published on: November 14, 2017

9.8K
Identification of RNA Fragments Resulting from Enzymatic Degradation using MALDI-TOF Mass Spectrometry
09:20

Identification of RNA Fragments Resulting from Enzymatic Degradation using MALDI-TOF Mass Spectrometry

Published on: April 11, 2022

3.8K

Area of Science:

  • Biochemistry
  • Analytical Chemistry
  • Molecular Biology

Background:

  • Accurate quantification of nucleic acids is crucial for molecular biology and diagnostics.
  • Existing methods may lack traceability or precision for absolute quantification.

Purpose of the Study:

  • To develop and validate a method for accurate, traceable quantification of RNA and DNA oligomers.
  • To establish a workflow combining acid hydrolysis with isotope dilution mass spectrometry (ID-MS).

Main Methods:

  • Oligonucleotides were hydrolyzed using formic acid to release nucleobases.
  • Nucleobases were quantified using liquid chromatography-tandem mass spectrometry (LC-MS/MS) with multiple reaction monitoring (MRM).
  • A novel cation-exchange separation with a pH gradient was employed for nucleobase resolution.

Main Results:

  • Formic acid hydrolysis achieved stoichiometric release of nucleobases from intact oligonucleotides.
  • The ID-LC-MS/MS method demonstrated excellent quantitative selectivity and accuracy.
  • Results showed good agreement with NIST Standard Reference Materials (SRMs) and other quantification techniques like ddPCR.

Conclusions:

  • Acid hydrolysis coupled with ID-LC-MS/MS provides a precise and traceable method for absolute quantification of RNA and DNA.
  • This approach is versatile and can be applied to quantify modified nucleobases and complex mixtures.