![]() This method allows easy penetration of the needle into the worm. When the tip is broken, the vacuole will leak out. Move the RWD Micromanipulatorto smash the needle against the edge of the glass slide. Place the coverslip on the agar plate filled with stock solution. After several minutes, observe the tip to see if there are any air bubbles. Use a small and thin chemical dropper to introduce the stock solution into the pipette through the tip of the needles. The purity of stock solution is one of the important factors affecting injection efficiency. For example, DNA, RNA, proteins and other substances. Step 3: DNA preparation and fill a needle-loading pipetteĪccording to the experimental requirements, choose the substances for stock solution. In general, the tip diameters are down to 1μm and the cone length is around 5~7mm. As such, the air bubbles can be discharged. It is recommended to use RWD capillary glass tube for pipette pulling to fill up the tip of the needles quickly with stock solution. RWD micropipette puller helps with generating two needles with stable and consistent performance. Remove the coverslip after the agarose solidifies. Place another coverslip on the agarose drop and gently press it to make it flattened. Drop 50ul of 2% hot agarose solution in the middle of the coverslip C. Mount the coverslip on a clean working stage with 2 tapes. Stack the plates for later use.įigure: Preparation of the agar plate. Let the agar plate dry overnight at room temperature or bake it at 80℃ for 1 hour. After the flattened agarose solidifies (~5 min), you can remove the coverslip on top by gently sliding. elegans Microinjetion Protocol Step 1: Preparation of agar platesĭrop 50ul of 2% hot agarose solution on a 24 × 60 mm glass coverslip and gently put another coverslip on top (beware of the air bubbles). elegans Microinjection as shown in the figure C. It is widely used in studying gene expression, gene function and genetic interactions inside the body of c elegans. elegans microinjection is the core technique in the C. Microinjection technology provides a capillary glass tube with a tip diameter of millimeters for the injection of nucleic acids into the worm’s gonad that is subsequently absorbed by the egg cells and generated into the form of extrachromosomal arrays. It can help us to study the mechanism of action from molecular and cellular level to system biological level in regard to the corresponding life cycle. elegans) is an important model organism used for studying animal genetics, ontogeny, and ethology. ![]()
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