DNA and genes
Genotyping, breeding of genetically modified lines, CRISPR genome editing, mutagenesis and cloning.
- Chromatin immunoprecipitation (ChIP-qPCR)Chromatin immunoprecipitation to test whether the transcription factor CREB, or an activity-dependent histone acetylation, is present at the promoter of the Fos gene and increases when neurons are stimulated.Guided experiment
- Restriction cloning of a gene into a plasmidConstruction of a recombinant plasmid carrying a fluorescent-protein gene, from the choice of vector and restriction enzymes to a sequence-verified clone.Guided experiment
- Breeding and genotyping a mouse lineBreeding and genotyping of one litter of a knockout mouse line, from the choice of breeders to a genotyped cohort.Guided experiment
- Gene knockout in cultured cells with CRISPR–Cas9Knockout of a gene in a cultured cell line with CRISPR–Cas9, from the choice of guide RNA to a clone that lacks the protein.Guided experiment
- Changing one amino acid: site-directed mutagenesisOne defined point mutation is made in a gene carried on a standard laboratory plasmid, from the choice of the amino-acid change to its confirmation in a sequencing trace.Guided experiment
- Mouse genotyping by PCR and gel electrophoresisGenotyping of a mouse litter for a knockout or floxed allele by PCR and agarose gel electrophoresis, with blind scoring and a test against Mendelian expectation.Simulator
- Breeding a conditional knockout lineA Cre–loxP breeding programme planned week by week, from founder crosses to an age-matched cohort with littermate controls, with a count of the surplus animals.Simulator
- CRISPR–Cas9 genome editingDesign of SpCas9 guides against a simulated gene, editing of cells or mouse zygotes, and validation of knockouts and point mutations by T7E1 assay and amplicon sequencing.Simulator
- DNA mutation and site-directed mutagenesisTranslation of a cloned gene, classification of mutations, and a defined point mutation taken through primer design, PCR, DpnI treatment, transformation and sequence-verified screening.Simulator
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