High-efficiency full-length cDNA cloning by biotinylated CAP trapper

Genomics
P CarninciC Schneider

Abstract

We have devised a method for efficiently constructing high-content full-length cDNA libraries based on chemical introduction of a biotin group into the diol residue of the cap structure of eukaryotic mRNA, followed by RNase I treatment to select full-length cDNA. The selection occurs by trapping the biotin residue at the cap sites using streptavidin-coated magnetic beads, thus eliminating incompletely synthesized cDNAs. When this method was used to construct a mouse brain full-length cDNA library, our evaluation showed that more than 95% of the total clones were of full length, and recombinant clones could be produced with high efficiency (1.2 x 10(7)/10 micrograms starting mRNA). The analysis of 120 randomly picked clones indicates an unbiased representation of the starting mRNA population.

Citations

Dec 10, 2002·Journal of Comparative Physiology. A, Neuroethology, Sensory, Neural, and Behavioral Physiology·E D JarvisS Lin
Apr 21, 2009·Nature Genetics·Ryan J TaftJohn S Mattick
Jun 24, 2005·Nature Methods·Matthias Harbers, Piero Carninci
May 20, 2008·DNA Research : an International Journal for Rapid Publication of Reports on Genes and Genomes·Mio OshikawaSeishi Kato
Aug 9, 2008·DNA Research : an International Journal for Rapid Publication of Reports on Genes and Genomes·Tingting LuBin Han
Oct 18, 2008·DNA Research : an International Journal for Rapid Publication of Reports on Genes and Genomes·Taishi UmezawaKazuo Shinozaki
Jan 20, 2009·DNA Research : an International Journal for Rapid Publication of Reports on Genes and Genomes·Kazuhiro SatoYuji Kohara
Jun 26, 2003·Nucleic Acids Research·Anason S HaleesZhiping Weng
Dec 31, 2005·Nucleic Acids Research·Riu YamashitaSumio Sugano
Jan 6, 2004·Nucleic Acids Research·Christian ClepetMichel Caboche
Feb 20, 2004·Nucleic Acids Research·Pavel A ZhulidovDmitry A Shagin
Nov 10, 2013·Plant & Cell Physiology·Keiichi Mochida, Kazuo Shinozaki
Jun 24, 2003·Genome Research·Piero CarninciYoshihide Hayashizaki
Dec 24, 2010·Genome Research·Roger A HoskinsSusan E Celniker
Sep 2, 2004·Cold Spring Harbor Symposia on Quantitative Biology·Y Hayashizaki
Oct 27, 2006·BMC Bioinformatics·Coral del ValStefan Wiemann
Jun 17, 2004·BMC Genomics·Ruth WellenreutherUNKNOWN German cDNA Consortium
Mar 17, 2009·BMC Biology·Naruo Nikoh, Atsushi Nakabachi
Mar 2, 2006·Genome Biology·Alistair R R ForrestSean M Grimmond
Sep 8, 2006·RNA·Stephen D BairdMartin Holcik
May 26, 2011·RNA·Carsten A RaabeTimofey S Rozhdestvensky
Aug 21, 2008·PloS One·Charles PlessyPiero Carninci
May 3, 2011·PloS One·Christian Clepet

❮ 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.