PMID: 9421498Feb 28, 1998Paper

Sequencing RNA by a combination of exonuclease digestion and uridine specific chemical cleavage using MALDI-TOF

Nucleic Acids Research
D A Tolson, N H Nicholson

Abstract

The determination of DNA sequences by partial exonuclease digestion followed by Matrix-Assisted Laser Desorption Time of Flight Mass Spectrometry (MALDI-TOF) is a well established method. When the same procedure is applied to RNA, difficulties arise due to the small (1 Da) mass difference between the nucleotides U and C, which makes unambiguous assignment difficult using a MALDI-TOF instrument. Here we report our experiences with sequence specific endonucleases and chemical methods followed by MALDI-TOF to resolve these sequence ambiguities. We have found chemical methods superior to endonucleases both in terms of correct specificity and extent of sequence coverage. This methodology can be used in combination with exonuclease digestion to rapidly assign RNA sequences.

References

Apr 1, 1979·Proceedings of the National Academy of Sciences of the United States of America·D A Peattie
Aug 1, 1977·Nucleic Acids Research·H Donis-KellerW Gilbert
Nov 25, 1987·Nucleic Acids Research·C EhresmannB Ehresmann
Jan 1, 1989·Methods in Enzymology·C S HahnJ H Strauss
Jan 1, 1989·Methods in Enzymology·Y Kuchino, S Nishimura
Jan 1, 1996·Mass Spectrometry Reviews·E NordhoffP Roepstorff

❮ Previous
Next ❯

Citations

Apr 1, 2003·Mass Spectrometry Reviews·Jörg Tost, Ivo G Gut
Sep 29, 2011·Nucleic Acids Research·John J TurnerSven Klussmann
Mar 26, 2003·Nucleic Acids Research·Stefan KrebsMartin Förster
May 10, 2012·Journal of Proteomics·Dominique RichterEduard F Stange
Oct 27, 2015·Journal of the American Chemical Society·Anders BjörkbomJack W Szostak
May 13, 2008·Rapid Communications in Mass Spectrometry : RCM·Yan ZouJerome Hochman
Feb 18, 2006·Trends in Biochemical Sciences·Benjamin Thomas, Alexandre V Akoulitchev
Aug 7, 2001·The Neuroscientist : a Review Journal Bringing Neurobiology, Neurology and Psychiatry·T Bonk, A Humeny
Apr 23, 2008·Current Protocols in Nucleic Acid Chemistry·C W Chou, P A Limbach
Apr 15, 2020·Molecular Omics·Richard Lauman, Benjamin A Garcia

❮ Previous
Next ❯

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.