ArticleScientific reports2019
A simple genotyping method to detect small CRISPR-Cas9 induced indels by agarose gel electrophoresis.
Article in Scientific reports, 2019. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 32 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
32 citing papers in PubMed.
- Voltage-gated calcium channels generate blastema Ca2+ fluxes restraining zebrafish fin regenerative outgrowth.Development (Cambridge, England) · 2025Article
- Same rule, different genes:Science advances · 2024Article
- Nesprin-2 coordinates opposing microtubule motors during nuclear migration in neurons.The Journal of cell biology · 2024Article
- Phenotypic correlates between clock genes and phenology among populations of Diederik cuckoo,Ecology and evolution · 2024Article
- A simple and effective genotyping workflow for rapid detection of CRISPR genome editing.American journal of physiology. Gastrointestinal and liver physiology · 2024Article
- Lymphocyte-Specific Protein 1 Regulates Expression and Stability of Endothelial Nitric Oxide Synthase.Biomolecules · 2024Article
- Optimising Electroporation Condition for CRISPR/Cas-Mediated Knockout in Zona-Intact Buffalo Zygotes.Animals : an open access journal from MDPI · 2023Article
- A histone demethylase links the loss of plasticity to nongenetic inheritance and morphological change.Nature communications · 2023Article
- Generation and initial characterization of mice lacking full-length BAI3 (ADGRB3) expression.Basic & clinical pharmacology & toxicology · 2023Article
- Genotyping MUltiplexed-Sequencing of CRISPR-Localized Editing (GMUSCLE): An Experimental and Computational Approach for Analyzing CRISPR-Edited Cells.The CRISPR journal · 2023Article
- Argonaute 5-mediated antiviral defense and viral counter-defense in Nicotiana benthamiana.Virus research · 2023Article
- Article
- PANTOTHENATE KINASE4, LOSS OF GDU2, and TRANSPOSON PROTEIN1 affect the canalization of tomato fruit metabolism.Plant physiology · 2023Article
- Genome editing of the vermilion locus generates a visible eye color marker for Oncopeltus fasciatus.Scientific reports · 2023Article
- Uncertainty Estimation for Quantitative Agarose Gel Electrophoresis of Nucleic Acids.Sensors (Basel, Switzerland) · 2023Article
- Infertility control of transgenic fluorescent zebrafish with targeted mutagenesis of theFrontiers in genetics · 2023Article
- Polymorphisms in exon 11 of the mptl-1 gene and monepantel resistance in Haemonchus contortus.Parasitology research · 2022Article
- One-Step Genotyping Method in loxP-Based Conditional Knockout Mice Generated by CRISPR-Cas9 Technology.Molecular biotechnology · 2022Article
- PPAR agonists attenuate lenalidomide's anti-myeloma activity in vitro and in vivo.Cancer letters · 2022Article
- Conventional Gel Electrophoresis-Resolvable Insertion/Deletion Markers for Individual Identification and Analysis of Population Genetics in Red-Crowned Cranes in Eastern Hokkaido, Japan.Animals : an open access journal from MDPI · 2022Article
Corrections and comments
PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.
Authors and funding
2 authors.
Funding
Abstract
CRISPR gene editing creates indels in targeted genes that are detected by genotyping. Separating PCR products generated from wild-type versus mutant alleles with small indels based on size is beyond the resolution capacity of regular agarose gel electrophoresis. To overcome this limitation, we developed a simple genotyping method that exploits the differential electrophoretic mobility of homoduplex versus heteroduplex DNA hybrids in high concentration agarose gels. First, the CRISPR target region is PCR amplified and homo- and hetero-duplexed amplicons formed during the last annealing cycle are separated by 4-6% agarose gel electrophoresis. WT/mutant heteroduplexes migrate more slowly and are distinguished from WT or mutant homoduplexes. Heterozygous alleles are immediately identified as they produce two distinct bands, while homozygous wild-type or mutant alleles yield a single band. To discriminate the latter, equal amounts of PCR products of homozygous samples are mixed with wild-type control samples, subjected to one denaturation/renaturation cycle and products are electrophoresed again. Samples from homozygous mutant alleles now produce two bands, while those from wild-type alleles yield single bands. This method is simple, fast and inexpensive and can identify indels >2 bp. in size in founder pups and genotype offspring in established transgenic mice colonies.
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.