ArticleNucleic acids research2002
Efficient isolation of targeted Caenorhabditis elegans deletion strains using highly thermostable restriction endonucleases and PCR.
Article in Nucleic acids research, 2002. The graph could read no effect estimate from its abstract, so it casts no vote on the map. An erratum has been issued. Cited by 16 papers.
What it found
Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.
The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.
The trial behind it
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.
Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.
Who cites it
16 citing papers in PubMed.
- Melatonin promotes sleep by activating the BK channel inProceedings of the National Academy of Sciences of the United States of America · 2020Article
- HRPU-2, a Homolog of Mammalian hnRNP U, Regulates Synaptic Transmission by Controlling the Expression of SLO-2 Potassium Channel inThe Journal of neuroscience : the official journal of the Society for Neuroscience · 2018Article
- SLO potassium channels antagonize premature decision making inCommunications biology · 2018Article
- BKIP-1, an auxiliary subunit critical to SLO-1 function, inhibits SLO-2 potassium channel in vivo.Scientific reports · 2017Article
- Mutations in KCNQ potassium channels cause pharyngeal pumping defects inmicroPublication biology · 2016Article
- Genomic Scars Generated by Polymerase Theta Reveal the Versatile Mechanism of Alternative End-Joining.PLoS genetics · 2016Article
- Restriction Cascade Exponential Amplification (RCEA) assay with an attomolar detection limit: a novel, highly specific, isothermal alternative to qPCR.Scientific reports · 2015Article
- SLO-2 potassium channel is an important regulator of neurotransmitter release in Caenorhabditis elegans.Nature communications · 2014Article
- Forward and reverse mutagenesis in C. elegans.WormBook : the online review of C. elegans biology · 2014Review
- large-scale screening for targeted knockouts in the Caenorhabditis elegans genome.G3 (Bethesda, Md.) · 2012Article
- An introduction to worm lab: from culturing worms to mutagenesis.Journal of visualized experiments : JoVE · 2011Article
- A semi-automated high-throughput approach to the generation of transposon insertion mutants in the nematode Caenorhabditis elegans.Nucleic acids research · 2007Article
- The phosphoinositide kinase PIKfyve/Fab1p regulates terminal lysosome maturation in Caenorhabditis elegans.Molecular biology of the cell · 2006Article
- Isolation of C. elegans deletion mutants following ENU mutagenesis and thermostable restriction enzyme PCR screening.Molecular biotechnology · 2006Article
- Discovery of a novel restriction endonuclease by genome comparison and application of a wheat-germ-based cell-free translation assay: PabI (5'-GTA/C) from the hyperthermophilic archaeon Pyrococcus abyssi.Nucleic acids research · 2005Article
- Dissection of K+ currents in Caenorhabditis elegans muscle cells by genetics and RNA interference.Proceedings of the National Academy of Sciences of the United States of America · 2003Article
Corrections and comments
- Erratum issued
Authors and funding
7 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Reverse genetic approaches to understanding gene function would be greatly facilitated by increasing the efficiency of methods for isolating mutants without the reliance on a predicted phenotype. Established PCR-based methods of isolating deletion mutants are widely used for this purpose in Caenorhabditis elegans. However, these methods are inefficient at isolating small deletions. We report here a novel modification of PCR-based methods, employing thermostable restriction enzymes to block the synthesis of wild-type PCR product, so that only the deletion PCR product is amplified. This modification greatly increases the efficiency of isolating small targeted deletions in C.elegans. Using this method six new deletion strains were isolated from a small screen of approximately 400 000 haploid genomes, most with deletions <1.0 kb. Greater PCR detection sensitivity by this modification permitted approximately 10-fold greater pooling of DNA samples, reducing the effort and reagents required for screens. In addition, effective suppression of non-specific amplification allowed multiplexing with several independent primer pairs. The increased efficiency of this technique makes it more practical for small laboratories to undertake gene knock-out screens.
Indexed as
Identifiers
What OpenQuestion holds
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.