C. elegans Gene Knockout Protocol The procedure is to mutagenize a large population of worms with trimethylpsoralen and UV irradiation, set up 1152 subpopulations, screen DNA made from this library for deletions in specific genes by nested PCR, and then to recover single worms carrying the deletions through a sib-selection process.
C. elegans RNAi Protocol Protocol for C. elegans RNAi. Includes: Transformation of competent cells; Blunt-end ligation; Preparation of competent cells; Dephosphorylation of linear plasmid DNA; Restriction Digest: EcoRV; Insert amplification from gDNA; Gel purification: QiaQuick gel purification kit; Mini-prep; Transformant Screening; Bacterial preparation and induction; Preparation of worms for RNAi feeding.
Cosuppression in C. elegans Protocol The cosuppression effect in C. elegans does not spread throughout the animal. Cosuppression in C. elegans can be triggered by highly repetitive transgenes that contain gene constructs.
Guide to Genetic Mapping in C. elegans A guide to genetic mapping in C. elegans. Includes: Introduction and basics; Two-point mapping; Three-point mapping; Deficiency and duplication mapping; Basics of SNP mapping; Mapping dominant mutations; Mapping suppressor/enhance mutations; Mapping of synthetic mutations.
Introduction of Double-Stranded RNA in C. elegans by Feeding Protocol Protocol describes an easily scalable way of introducing double-stranded RNA (dsRNA) in Caenorhabditis elegans: feeding the nematode with bacteria that express dsRNA. When using an RNase-III-negative Escherichia coli strain (HT115), the efficiency of this method is comparable to the alternative.
Introduction of Double-Stranded RNA in C. elegans by Injection Protocol Double-stranded RNA (dsRNA) can be efficiently introduced into Caenorhabditis elegans by microinjection into the gonad, the gut, or the body fluid. The RNAi effect will spread within the nematode, exerting an effect beyond the site of injection.
Introduction of Double-Stranded RNA in C. elegans by Soaking Protocol Double-stranded RNA (dsRNA) can be introduced into Caenorhabditis elegans by soaking the animals in a solution of dsRNA. Alternative methods are dsRNA injection (see Introduction of Double-stranded RNA in C. elegans by Injection) and feeding the animals with bacteria that produce dsRNA.
