A working protocol
Got the electroporator to Djerassi Scientific Delirium residency... Considering using my sperm... And some centipedes" DNA... Wondering if you have any sense as to a DIY sperm salt rinse media/protocol and refresh media... I mean like Gatorade grade materials... I'm way out in the mountains.
Sent from my iPadilla
To: Adam Zaretsky
Not sure how resilient semen is to washes with water but all you would need is a centrifuge, some distilled water, and glycerin. The glycerin can we purchased from a drug store like Duane Reade. Glycerin and glycerol are equivalent. The idea is to NOT make the fluid conductive so you would need to rid yourself of all the salts in the solution.
Spindown, discard the liquid and resuspend the pellet with water until its fully dissolved in solution again
then spin down again and discard the pellet and resuspend in cold distilled water
Spin down once more and this time resuspend the pellete in 10% glycerol (9 parts water to one part glycerin) also ice cold.
Then take 50uL per reaction. The centipede DNA can be isolated potentially by grinding and boiling the centipede mash in a water salt and tiny drop of dish soap (like tiniest drop) for about 5-10 minutes. Just nuke a mug of water until rolling boil and then let the sample sit in the mug for 10mims. Then spin down the tube and KEEP the liquid phase while discarding the solid pellet. Move this liquid to a new tube. Add equal volume everclear vodka as there is sample, mix, and spin down for 10mins at max speed. That should give you a crude DNA isolate. Remove vodka
Let air dry
Resuspend 100 microlites DH2O (distilled water) (pellet may be invisible, pipette up and down)
Add 10uL of the centipede dna to the sperm sample in the cuvette like we did before and zap the bastards at around 1800v. Good luck and godspeed!
Sebastian S. Cocioba CEO & Founder New York Botanics, LLC Blog: ATinyGreenCell.com