Today was a great day. I planted the sterilized cuttings in the jars, and it all went pretty well. I cut off four of the developing leaves of my D. burmannii (hopefully it isn't damaged that badly), washed them in hot soap and water, and proceeded with the following procedure. In each of the steps I shook the jar vigorously.
INSIDE HOOD
1. Dunk explants in 91% alcohol for 10 seconds. Skip this for seeds.
2. Dunk in 3% H2O2 for 4 minutes.
3. Dunk in 10% bleach solution for 7 minutes. Put seeds in here for 4 minutes.
4. Place in sterile jar of water and rinse for 5 minutes. Repeat.
5. Pull samples out of jar with sterilized forceps.
6. Loosen cap on TC jar.
7. Place explants in jar extremely quickly.
STERILIZATION:
1. Using tiny spray bottle, spray 10% bleach solution on every surface.
2. Generously spray jars with 10% bleach solution.
3. Wash hands with soap and water, then wipe with 70% alcohol.
To prepare the steril water, I filled loose jars with distilled water and pressure cooked them for 25 minutes. For the bleach solution, I used 2/3 cup of extra-concentrated bleach in 9 1/3 cups of tap water. I figured any impurities in the tap water would be washed off in the final rinse. If I were to do this again, I would use flat forceps the whole time, for this I used surgical forceps with teeth that I am sure damaged the cuttings. I used flat forceps for the final transfer, because I did not want the cuttings to stick.
Here are some pictures from today:
I was about to try and sterilize forceps with a lighter, then realized I had these covering every surface. An oxygen source, two potent fuels, and a reactive poisonous vapor source.
<a href="http://www.flickr.com/photos/84442298@N03/8583529477/" title="DSC02237 by Sundrew, on Flickr"><img src="http://farm9.staticflickr.com/8098/8583529477_03c2ac75c1_z.jpg" width="640" height="480" alt="DSC02237"></a>
The aftermath.
<a href="http://www.flickr.com/photos/84442298@N03/8583529479/" title="DSC02238 by Sundrew, on Flickr"><img src="http://farm9.staticflickr.com/8367/8583529479_9c759b1dcd_z.jpg" width="640" height="379" alt="DSC02238"></a>
The river of bleach.
<a href="http://www.flickr.com/photos/84442298@N03/8583529437/" title="DSC02239 by Sundrew, on Flickr"><img src="http://farm9.staticflickr.com/8518/8583529437_5bce642aa9_z.jpg" width="640" height="480" alt="DSC02239"></a>
The best cutting.
<a href="http://www.flickr.com/photos/84442298@N03/8584629154/" title="DSC02241 by Sundrew, on Flickr"><img src="http://farm9.staticflickr.com/8243/8584629154_98e44d3917_z.jpg" width="640" height="480" alt="DSC02241"></a>
The cutting with terrible aim.
<a href="http://www.flickr.com/photos/84442298@N03/8583528617/" title="DSC02244 by Sundrew, on Flickr"><img src="http://farm9.staticflickr.com/8380/8583528617_f15b9008c7_z.jpg" width="640" height="480" alt="DSC02244"></a>
The jars' new home (I realized this was close to the lights and moved them to my basement's stone floor where they will stay in the 60s)!
<a href="http://www.flickr.com/photos/84442298@N03/8583528537/" title="DSC02243 by Sundrew, on Flickr"><img src="http://farm9.staticflickr.com/8371/8583528537_8b589d0a9c_z.jpg" width="640" height="480" alt="DSC02243"></a>