Post by mrflytrap2 on Jan 10, 2008 5:39:31 GMT
Well I finished round 2 of my tissue cultures yesterday. Lessons learned from round one that I put into this one.
-Don't do tissue culture. Hehe, I opted to do everything off of seeds, it was the only thing that was a success last time and now have some Capensis Alba seeds growing on my plant rack. When I did my first round, I realized how many procedures were new to me, how many things I had to figure out to do 'just right' yet. This time around I did 3 packet of seeds that I got from the ICPS. I did this instead of TC as a way to learn how to work with the jars, work with the seeds, not explode the media in the microwave, keeping everything clean, learn how to measure accurate, etc.
- Purchased a Digital pH meter, doing things by paper last time was a horrible experience, I couldn't tell if I was getting closer, further, or doing anything at all. I saved some of my old media and tested it this round, turns out I wasn't that far off! But still, it added a lot of delay and frustration to the process that I didn't need at the time.
- Constructed a Flow Hood, I realized that the places where I could work are horrible, they are dusty, drafty, and dirty. Even then though, I have jars now over 6 months old that were unused but with sterile media in them that are still contaminate free. Where I placed them for growing though was a mistake as it was too close to my other plants with extra moisture that could splash onto the jars if I wasn't careful. (I wasn't) The LFH should help for this situation.
- Numbered each jar, & took notes on different batches, things I think I did wrong, etc. For some, I still couldn't multi-task very well, and if I fumbled things around, I noted it after. Also I tried a few different techniques for getting small seeds into the jars. Each method was noted by the jar number as well as how I sterilized the media in the microwave. At least I'll have some notes to review on the first contaminated looser that shows up.
Things learned that will be used in round 3:
Already I have a few things I want to try for round 3...
- Glass pipettes, the plastic pipettes are pretty much useless in my opinion, or at least I can't use them very well. I'm not confident that I was able to clean them properly from the previous round, or could even measure accurately with them. I already purchased a pipette pump and disposable 2mL glass pipettes. As a test I tried to measure out 2mL of liquid with the plastic, and remeasure it with the glass. I found out that I really only got 1.4mL of liquid on my first measure with the plastic pipettes.
- Working with small things. Even in this round I still fumbled around with trying to extract seeds from washing them and putting them into jars. I'm gonna try to do some more research into this and I noted a few methods that I tried. I've been thinking of trying to do some practice runs instead with pepper to try and simulate 'seed moving training.' ( or some other solid spice )
I hope this helps some of you, I know I was borderline with a few of these things on my first round ( like the pH meter) maybe it will help some with choices or things to look out for. Hopefully I'll have some pics to share in a few weeks of baby fly traps, binata's and capensis.
Thanks!
Nathan
(I'm posting this on 2 forums, sorry if it's a repeat!)
-Don't do tissue culture. Hehe, I opted to do everything off of seeds, it was the only thing that was a success last time and now have some Capensis Alba seeds growing on my plant rack. When I did my first round, I realized how many procedures were new to me, how many things I had to figure out to do 'just right' yet. This time around I did 3 packet of seeds that I got from the ICPS. I did this instead of TC as a way to learn how to work with the jars, work with the seeds, not explode the media in the microwave, keeping everything clean, learn how to measure accurate, etc.
- Purchased a Digital pH meter, doing things by paper last time was a horrible experience, I couldn't tell if I was getting closer, further, or doing anything at all. I saved some of my old media and tested it this round, turns out I wasn't that far off! But still, it added a lot of delay and frustration to the process that I didn't need at the time.
- Constructed a Flow Hood, I realized that the places where I could work are horrible, they are dusty, drafty, and dirty. Even then though, I have jars now over 6 months old that were unused but with sterile media in them that are still contaminate free. Where I placed them for growing though was a mistake as it was too close to my other plants with extra moisture that could splash onto the jars if I wasn't careful. (I wasn't) The LFH should help for this situation.
- Numbered each jar, & took notes on different batches, things I think I did wrong, etc. For some, I still couldn't multi-task very well, and if I fumbled things around, I noted it after. Also I tried a few different techniques for getting small seeds into the jars. Each method was noted by the jar number as well as how I sterilized the media in the microwave. At least I'll have some notes to review on the first contaminated looser that shows up.
Things learned that will be used in round 3:
Already I have a few things I want to try for round 3...
- Glass pipettes, the plastic pipettes are pretty much useless in my opinion, or at least I can't use them very well. I'm not confident that I was able to clean them properly from the previous round, or could even measure accurately with them. I already purchased a pipette pump and disposable 2mL glass pipettes. As a test I tried to measure out 2mL of liquid with the plastic, and remeasure it with the glass. I found out that I really only got 1.4mL of liquid on my first measure with the plastic pipettes.
- Working with small things. Even in this round I still fumbled around with trying to extract seeds from washing them and putting them into jars. I'm gonna try to do some more research into this and I noted a few methods that I tried. I've been thinking of trying to do some practice runs instead with pepper to try and simulate 'seed moving training.' ( or some other solid spice )
I hope this helps some of you, I know I was borderline with a few of these things on my first round ( like the pH meter) maybe it will help some with choices or things to look out for. Hopefully I'll have some pics to share in a few weeks of baby fly traps, binata's and capensis.
Thanks!
Nathan
(I'm posting this on 2 forums, sorry if it's a repeat!)
