Well I had been itching to try it for a while so I read up on several different people’s ideas and combined some with a few bright ideas of my own. Mostly I'm flying by the seat of my pants but hey! I am learning!
The medium I used was based on one of C.D Moulton's from his book working with cornstarch. I think I altered the calcium amount added to the medium a bit because I couldn't find a Calcium Citrate solution. I had to make my own.
I made a glove box out of plexi glass sheets that I bought at home depot. 3 solid sides and the top are held on by hinges. Then I glued a rubber seal all around the top of the opening, so when I close the lid there is no gap for air to pass through. It fits nice and snug. In the front, I cut holes and glued long thick dishwasher type gloves.
I used glass lab specimen jars for mother flasks (ordered from medical supply warehouse). They have plastic screw on tops. I soaked both the tops and the jars and one turkey baster in a giant bucket of 10% bleach solution for 24 hours.
The next day, while the jars, tops, and turkey baster are still soaking in the bucket, I mixed up my flask medium. I also cut 4 pieces of tinfoil per flask to go over the top of the jars once I filled them with medium.
I put on rubber disposable gloves, and then remove the turkey baster from the bucket. Suck up some medium squirt it into each jar one by one (careful so as not to touch the sides of the jar top with anything especially the medium). As I filled each jar, I quickly covered them with 4 pieces of the tinfoil apiece. When I had finished filling my jars, I loaded them into my pressure cooker. It contained 2 inches of water on the bottom. I locked it and cooked at 15 psi for 10 mins.
While it was cooling off, I sprayed the inside of my glove box with 10% bleach solution that I put into a spray bottle. Waited 5 mins and then began putting in my tools.
I used 1 large long tray 5 inches deep that I fill with 10% bleach solution. Inside it, I placed 20 still capped sterile syringes (with needles) that I got from a veterinary supply warehouse. 1 metal lab shovel (I dono how else to describe this tool. It looks like a tiny silver shovel) and 20 test tubes with plastic caps. (caps removed and floating with everything in the tray)
Against the back wall of the glove box, I put 1 squirt bottle container of distilled water, My container with seeds, and one wide mouth container of 10% bleach solution with nothing in it. One clean container filled with 20 cotton swabs (not synthetic). A black magic marker (for writing on jars). And one small empty tub which I called my *trash* tub. Then I took the jar tops (which were still floating in the bucket, and I put them in the case. Lastly, I added one spray bottle with 10% bleach solution into the glove box. Then I sprayed all my tools and glove box down again inside the glove box with the bleach solution. Waited another 5 mins.
When I did it, I used 2 spray bottles so I could keep one inside the glove box at all times (just in case)
By then my jars covered in tinfoil were cool and the pressure valve released so I put all my jars into my glove box. Then I sprayed everything again with the 10% bleach and shut the top of the case. This time I waited 10 mins.
I took one test tube at a time filled it with seed, poured 10% bleach solution into it capped it and shook it, let it rest, shook it, let it rest for 5 mins. Then I decanted some water into my empty *trash tub* and did it again. Then I added some of the distilled water from the squirt bottle into the test tube and did the same thing. When I was finished I decanted most of the water off save for an inch of seed and water. Then I grabbed a syringe floating in the long tray, popped the cap off and sucked up some seed water. Then grab a cotton swab that has been saturated in 10% bleach solution (from all the spraying) and I used it to wrap around the needle on the syringe and I hold it in place so no part of the needle is exposed. Then I punctured the tinfoil with the syringe making the smallest of holes with the needle. (I never remove or touched the tinfoil once it comes out of the cooker; likewise, the cotton saturated in bleach solution that covers the needle is the only thing that touches the tinfoil or the hole. It should I think kill any bacteria near the pinhole if there is any) Squirt in the seeds then I quickly removed the syringe, and throw it and the used cotton swab into the empty *trash* tub and cap the jar with its plastic lid.
I did it repeatedly, each time using a new syringe and a new swab.
They do the same thing when they take blood or give you a shot at the doctor. So I figure it should work hehe. I dono, part of my method I just made up on my own because I don't know anyone who does this. I'm hoping it works!
Regardless, I am keeping records of each thing I do and if this time fails, I will try again changing my approach a little bit each time until it does work.
DJ