Injection of dsRNA and Electroporation in Avian Embryos Protocol Protocol describes a method for in ovo transfection of avian embryos with double-stranded RNA (dsRNA). The dsRNA is injected into the spinal cord of the embryo. Subsequent electroporation facilitates the cellular uptake of the dsRNA molecules. It may be necessary to optimize the stage of the embryo and the electroporation procedure to improve the effectiveness of in ovo RNAi—cell competence changes with differentiation.
Production of dsRNA for RNAi in Avian Embryos Protocol Protocol describes the production of double-stranded RNA (dsRNA) from fragments of cDNAs of candidate genes. The cDNA fragments must be cloned in plasmids with a flanking SP6 and T7 promoter (e.g., pSP72 or pCRII). The plasmid is linearized and sense and antisense RNAs are produced separately by in vitro transcription.
Staining of Tissue Slices for Analysis of Axonal Pathfinding in dsRNA-Treated Avian Embryos Protocol describes a method to stain nerve fibers in tissue slices of avian embryos using an antibody against the 160-kD subunit of neurofilaments. This allows the comparison of the branching pattern of motor and sensory neurons between control and experimental embryos. The tissue is cut in slices using a vibratome or tissue slicer. The protocol is suitable for older embryos after approximately stage 33 and regions that are not accessible by whole-mount analysis.
Whole-Mount Preparations for Analysis of Axonal Pathfinding in dsRNA-Treated Avian Embryos Protocol describes a method for staining nerve fibers in whole-mount preparations of avian embryos using an antibody against the 160-kD subunit of neurofilaments. This allows the comparison of the branching pattern of motor and sensory neurons between control and experimental embryos. This protocol has been successfully applied for embryos at different stages up to about stage 33 (7 days of incubation).
C and M Law Corporation's
Los Angeles personal injury attorney firm has been serving the city’s residents for over 45 years. People who think they do not need the services of an experienced personal injury attorney, invariably find out the hard way that they should have chosen that right lawyer in the very beginning. Regardless of the type of accident or injury, we have the experience to successfully represent you and your family. If you or someone you know has been injured through the negligence or recklessness of others, come see us. Voted in the top one percent of trial lawyers in the USA, we can help get you the compensation you and your loved ones deserve. The personal injury attorney Los Angeles firm of C and M Law Corporation has won an excess of 2 Billion Dollars in settlements!