ArticleNucleic acids research2002
Systematic sequencing of cDNA clones using the transposon Tn5.
Article in Nucleic acids research, 2002. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 36 papers.
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Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.
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Who cites it
36 citing papers in PubMed, 59 citations in OpenAlex.
- Tagmentation-based analysis reveals the clonal behavior of CAR-T cells in association with lentivector integration sites.Molecular therapy oncolytics · 2023Article
- Enabling low-cost and robust essentiality studies with high-throughput transposon mutagenesis (HTTM).PloS one · 2023Article
- Elucidation of Essential Genes and Mutant Fitness during Adaptation toward Nitrogen Fixation Conditions in the Endophyte Azoarcus olearius BH72 Revealed by Tn-Seq.Microbiology spectrum · 2022Article
- Comprehensive understanding of Tn5 insertion preference improves transcription regulatory element identification.NAR genomics and bioinformatics · 2021Article
- Fast and inexpensive whole-genome sequencing library preparation from intact yeast cells.G3 (Bethesda, Md.) · 2021Article
- Transposon Insertion Sequencing, a Global Measure of Gene Function.Annual review of genetics · 2020Review
- Large-Scale Low-Cost NGS Library Preparation Using a Robust Tn5 Purification and Tagmentation Protocol.G3 (Bethesda, Md.) · 2018Article
- Large-Scale Transposition Mutagenesis of Streptomyces coelicolor Identifies Hundreds of Genes Influencing Antibiotic Biosynthesis.Applied and environmental microbiology · 2017Article
- The design and analysis of transposon insertion sequencing experiments.Nature reviews. Microbiology · 2016Review
- Mechanisms of DNA Transposition.Microbiology spectrum · 2015Review
- Target capture during Mos1 transposition.The Journal of biological chemistry · 2014Article
- Article
- Review
- Burkholderia cenocepacia conditional growth mutant library created by random promoter replacement of essential genes.MicrobiologyOpen · 2013Article
- Large-scale sequencing based on full-length-enriched cDNA libraries in pigs: contribution to annotation of the pig genome draft sequence.BMC genomics · 2012Article
- Transposon mutagenesis of the anaerobic commensal, Bacteroides fragilis, using the EZ::TN5 transposome.FEMS microbiology letters · 2012Article
- Insertion site preference of Mu, Tn5, and Tn7 transposons.Mobile DNA · 2012Article
- Target site selection by the mariner-like element, Mos1.Genetica · 2010Article
- Article
- Temporal control of message stability in the life cycle of double-stranded RNA bacteriophage phi8.Journal of virology · 2009Article
Corrections and comments
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Authors and funding
10 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
In parallel with the production of genomic sequence data, attention is being focused on the generation of comprehensive cDNA-sequence resources. Such efforts are increasingly emphasizing the production of high-accuracy sequence corresponding to the entire insert of cDNA clones, especially those presumed to reflect the full-length mRNA. The complete sequencing of cDNA clones on a large scale presents unique challenges because of the generally small, yet heterogeneous, sizes of the cloned inserts. We have developed a strategy for high-throughput sequencing of cDNA clones using the transposon Tn5. This approach has been tailored for implementation within an existing large-scale 'shotgun-style' sequencing program, although it could be readily adapted for use in virtually any sequencing environment. In addition, we have developed a modified version of our strategy that can be applied to cDNA clones with large cloning vectors, thereby overcoming a potential limitation of transposon-based approaches. Here we describe the details of our cDNA-sequencing pipeline, including a summary of the experience in sequencing more than 4200 cDNA clones to produce more than 8 million base pairs of high-accuracy cDNA sequence. These data provide both convincing evidence that the insertion of Tn5 into cDNA clones is sufficiently random for its effective use in large-scale cDNA sequencing as well as interesting insight about the sequence context preferred for insertion by Tn5.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.