PMID: 8938436Nov 1, 1996Paper

End sequence determination from large insert clones using energy transfer fluorescent primers

Genome Research
M MarraE R Mardis

Abstract

Genome mapping strategies depend heavily on confirmatory data of several types to establish overlaps between contiguous stretches of cloned DNA derived from genomic regions. One type of ancillary data that can contribute to establishing these overlaps is DNA sequence data derived from the ends of large (> 30 kb) inserts in genomic clones. This type of data can be difficult to obtain routinely, because large clones are often unstable and microgram quantities of highly purified DNA are required in each sequencing reaction to obtain sufficient signal for accurate base calling and maximum read length. Recently, we have been experimenting with methods to consistently obtain up to 800 bases of high-quality sequence data from the ends of large insert clones using ThermoSequenase DNA polymerase and Energy Transfer fluorescent primers. Our experimental approach and results, described in this paper, indicate that routinely obtaining high-quality sequence data from the ends of large insert genomic clones is feasible. Such data can contribute to the assessment of common regions between large insert clones, to the establishment of conservation of synteny between closely related species, and to the detection of additional contiguous clones.

References

Feb 25, 1990·Nucleic Acids Research·H VossW Ansorge
Jul 3, 1995·Proceedings of the National Academy of Sciences of the United States of America·S Tabor, C C Richardson
May 9, 1995·Proceedings of the National Academy of Sciences of the United States of America·J JuR A Mathies
Oct 1, 1986·Proceedings of the National Academy of Sciences of the United States of America·A CoulsonJ Karn

❮ Previous
Next ❯

Citations

Jul 15, 1997·Nucleic Acids Research·L G LeeJ M Cassel
Feb 12, 1998·Nucleic Acids Research·B B RosenblumS M Chen
Oct 12, 2011·Current Protocols in Molecular Biology·Barton E SlatkoFrederick M Ausubel
Oct 12, 2011·Current Protocols in Molecular Biology·Jay A ShendureBarton E Slatko
Dec 2, 1999·Nature Genetics·G T MarthW R Gish
Sep 10, 2005·Expert Review of Molecular Diagnostics·Xiaopeng BaiJingyue Ju
Jan 10, 1998·Genome Research·M A MarraR H Waterston

❮ 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.