Round A/Round B Amplification of DNA for Microarrays

Cold Spring Harbor Protocols
Oliver Rando

Abstract

The goal of this procedure is to randomly amplify a sample of DNA to achieve the best possible sequence representation for hybridization to microarrays. This protocol has been used successfully to amplify genomic representations starting with <10 ng of DNA. The protocol consists of three sets of enzymatic reactions. In Round A, Sequenase is used to extend randomly annealed primers to generate templates for subsequent PCR. During Round B, a specific primer is used to amplify the previously generated templates. Amplified material can be labeled by adding fluorescent moieties to the nucleic acids. Alternatively, Round C in this protocol can be used to incorporate either aminoallyl-dUTP or Cy-dye-coupled nucleotides during additional PCR cycles. This protocol may be unsuitable for amplifying material smaller than 250 bp because such material will not be amplified uniformly.

References

Aug 3, 2019·Cold Spring Harbor Protocols·Oliver Rando
Aug 3, 2019·Cold Spring Harbor Protocols·Oliver Rando
Aug 3, 2019·Cold Spring Harbor Protocols·Oliver Rando

❮ Previous
Next ❯

Citations

Jun 7, 2019·Cold Spring Harbor Protocols·Oliver Rando
Aug 3, 2019·Cold Spring Harbor Protocols·Oliver Rando
Aug 3, 2019·Cold Spring Harbor Protocols·Oliver Rando
Aug 3, 2019·Cold Spring Harbor Protocols·Oliver Rando

❮ 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.