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Related Concept Videos

Maxam-Gilbert Sequencing01:05

Maxam-Gilbert Sequencing

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.
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Sanger Sequencing

DNA sequencing is a fundamental technique that is routinely used in the biological sciences. This method can be applied to a range of questions at different scales - from the sequencing of a cloned DNA fragment or the study of a mutation in a gene up to whole-genome sequencing. However, despite the widespread use of sequencing today, it was not until 1977 that Fredrick Sanger and his collaborators developed the chain-termination method to decode DNA sequences. It relies on the separation of a...
Next-generation Sequencing03:00

Next-generation Sequencing

The first human genome sequencing project cost $2.7 billion and was declared complete in 2003, after 15 years of international cooperation and collaboration between several research teams and funding agencies. Today, with the advent of next-generation sequencing technologies, the cost and time of sequencing a human genome have dropped over 100 fold.
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Although all next-generation methods use different technologies, they all share a set of standard features.

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Related Experiment Video

Updated: May 29, 2026

2D-HELS MS Seq: A General LC-MS-Based Method for Direct and de novo Sequencing of RNA Mixtures with Different Nucleotide Modifications
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Methods for L-ribooligonucleotide sequence determination using LCMS.

John J Turner1, Johannes S Hoos, Stefan Vonhoff

  • 1NOXXON Pharma AG, Max-Dohrn-Strasse 8-10, D-10589 Berlin, Germany. jturner@noxxon.com

Nucleic Acids Research
|September 28, 2011
PubMed
Summary
This summary is machine-generated.

Verifying therapeutic oligonucleotide sequences is crucial. New methods enable accurate sequence determination for Spiegelmers, highly stable RNA enantiomers, using chemical degradation and liquid chromatography-mass spectrometry.

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Linear Amplification Mediated PCR – Localization of Genetic Elements and Characterization of Unknown Flanking DNA
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Linear Amplification Mediated PCR – Localization of Genetic Elements and Characterization of Unknown Flanking DNA
11:58

Linear Amplification Mediated PCR – Localization of Genetic Elements and Characterization of Unknown Flanking DNA

Published on: June 25, 2014

Area of Science:

  • Biochemistry
  • Drug Development
  • Analytical Chemistry

Background:

  • Nucleic acid-based therapeutics require sequence verification for drug development.
  • Therapeutic oligonucleotides possess high nuclease resistance, complicating sequence analysis.
  • Spiegelmers, enantiomers of RNA, offer enhanced enzymatic degradation resistance.

Purpose of the Study:

  • To develop and validate methods for determining the sequence of Spiegelmers.
  • To address the challenges in sequencing nuclease-resistant therapeutic oligonucleotides.

Main Methods:

  • Chemical degradation of Spiegelmers after 5'-end labeling.
  • Analysis of generated fragments using liquid chromatography-mass spectrometry (LC-MS).
  • Derivation of the complete Spiegelmer sequence from fragment data.

Main Results:

  • Successful sequence determination for two distinct Spiegelmers (NOX-E36 and NOX-A12).
  • Demonstration of method specificity using a mismatch control for NOX-E36.
  • Validation of the applicability of the developed sequencing approach.

Conclusions:

  • The described method enables reliable sequence determination of Spiegelmers.
  • This technique is essential for quality control in the development of Spiegelmer-based therapeutics.
  • The approach enhances the verification process for nuclease-resistant nucleic acid drugs.