RNAi Mouse Embryos and Oocytes Protocols

Collection of Early Mouse Embryos for RNAi Protocol Protocol describes a method to collect early embryos from 6-week-old mice.  Subsequently, the isolated embryos can be injected with double-stranded RNA to induce knockdown of a gene of interest.

Collection of Mouse Oocytes for RNAi Protocol Protocol describes a method to collect oocytes from 6-week-old mice.  Subsequently, the isolated oocytes can be injected with double-stranded RNA to induce knockdown of a gene of interest.

Microinjection of dsRNA into Mouse Oocytes and Early Embryos Protocol Protocol describes how to introduce a double-stranded RNA (dsRNA) of choice into mouse oocytes or fertilized one-cell embryos by microinjection.  For collection of mouse oocytes and early embryos, see Collection of Mouse Oocytes for RNAi and Collection of Early Mouse Embryos for RNAi.

Preparation of dsRNA Molecules for RNAi in Mouse Oocytes and Early Embryos Protocol Protocol describes in vitro transcription with the SP6 polymerase.  For other polymerases (T3, T7), simply use the appropriate buffer and desired RNA polymerase instead of the SP6 polymerase.

RNA Isolation and RT-PCR from dsRNA-Treated Mouse Oocytes and Early Embryos Protocol Protocol describes the use of RT-PCR to confirm knockdown of a gene of interest in RNAi-treated mouse oocytes and early embryos.  All of the solutions used during RNA isolation and reverse transcription must be Rnase-free