Electroporation Protocol, the making of centiSperm
Electroporation, the making of centiSperm
A – Mix sperm and DNA in the electroporation vial (cuvette)
For each reaction mix 10 microliters of centipede DNA and 50 microliters of desalinated sperm directly inside the electroporation cuvette
B - Put cuvette inside of the electroporator
C - Set voltage
I did one reaction at 200 volts which seemed standard in the sperm electroporation. I also ran a second reaction at 1800 volts just to make it rough. Sometimes when you are running a rough DIY lab its better to go for overkill than treading to closely to the protocol. More is sometimes a matter of affect. But more actually often does the trick!
D - Press button twice to fire machine. Wait for beep (about 2 seconds)
E – apply refresh media
For this lab I used 10% vitaminwater® XXX açai-blueberry- pomegranate flavored (Glaceau) put through a 3 micron Gelman Filter
The refresh media was prepared before the electroporation reaction.