To begin, use a pipette puller to stretch a 20 micrometer wide glass capillary. Place an extracted mouse uterus on a dry paper towel. Then, insert the tip of a pair of fine scissors into the myometrial side of the uterus, and slowly run the edge of the blade through the whole length and cut it.
Now, expose and cut the decidui. Transfer them into a dish with growth medium. Dissect the decidui to extract the embryos.
Transfer the embryos into a 60 millimeter Petri dish containing warm growth medium. With a pair of fine forceps, carefully cut the tail and the head of the embryo, and leave only the trunk of the body. Carefully remove as much tissue as possible from the trunk without damaging the heart or pulmonary and aorta arteries.
Insert a new stretched out needle into the tip of the mouth pipette, and carefully aspirate and load 10 microliters of a prepared plasmid mix into the stretched out needle. Place a single heart into a clean 60 millimeter Petri dish containing prewarmed PBS. Place the dish under a dissecting microscope.
Adjust the distance between the poles of positive and negative electrodes to about four millimeters. Next, use a needle in the mouth pipette to gently puncture the most superficial layer of the heart. Transfer one to two microliters of the plasmid mix into the heart.
Remove the needle carefully. Hold the electrode in place so that the heart is located between both poles. Then, electroporate the heart.
Transfer the electroporated heart to a new 12 well plate, containing one milliliter of growth medium. Incubate the plate at 37 degrees Celsius under 5%carbon dioxide until analysis.
