Electroporation Protocol, the making of centiSperm

By Adam Zaretsky

Electroporation, the making of centiSperm

 

Comic Rendition:  Matteo Farinella Matteo Farinella of http://matteofarinella.com

 

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

cuvette_centiSperm_4175

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

          centiSperm_4181

The refresh media was prepared before the electroporation reaction.    
 

Untitled-48 photo credit: David Bowen

 

                electroporator

     centiSperm as a new media is now available for experimentation.          IMG_1927_4378