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By Jessy2363
Posts:  4
Joined:  Thu Aug 13, 2020 5:26 am
#362672
Hi, I'm an absolute beginner with TC. I've spent the past week or so reading about it and trying to gather up everything I need to begin.
So I've read it's best to have a vent in the vessel to allow for gas exchange whilst obviously keeping the bad guys out. I'm thinking about using erlenmeyer flasks with a rubber/silicon bung in the top and drill a hole in it and stuff it densely with cotton wool. Will this work to keep the nasty microbes and fungi out? Has anyone else done this? I know I need something that can filter down to 0.2um or is that a bit over the top?
I want to be able to reuse my vessels over and over hence wanting to use pyrex glass or baby food jars or something that is glass. But this whole vent hole thing has be a bit bamboozled. Any ideas? please :)

I found some little round adhesive discs but they are very expensive. Like $1 each, and also I doubt they'd stick that good to a rubber bung although I'm unsure. Maybe I just bite the bullet and buy those. Any cool hacks any knows of in regards to vents in vessels that they wanna share?

I did a search on this forum and haven't managed to find a lot of information on this topic, unless I've missed it.
Jess
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By Matt
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Joined:  Mon Apr 21, 2008 11:28 pm
#362735
Jessy2363 wrote:So I've read it's best to have a vent in the vessel to allow for gas exchange whilst obviously keeping the bad guys out.
I have read this as well but have never done it and the plants I TC do fine. The risk of contamination is too great with everything I've tried when allowing air to flow.

Start simple and with the idea of keeping clean cultures. If they don't grow well for you without good air exchange, adjust then. But my guess is that they will grow fine even doubly sealed to keep the "nasties" out of the cultures.
By Jessy2363
Posts:  4
Joined:  Thu Aug 13, 2020 5:26 am
#362757
Thank you! yes it's possible I am overthinking it and becoming obsessional about the need to wanna vent the container. I literally spent hours on countless lab supplies websites looking for something that would filter down to 0.2um. Then I saw some already vented tissue culture jars with lids and they only filtered down to 0.4um so obviously I don't need to filter down to 0.2um. But yes you are right, maybe I should just try without the whole vent thing and see how it goes. I mean it's SO much easier finding vessels without vents.
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By Matt
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Posts:  21339
Joined:  Mon Apr 21, 2008 11:28 pm
#362826
Jessy2363 wrote:yes it's possible I am overthinking it and becoming obsessional about the need to wanna vent the container.
As a seasoned TC person, I would say that you are. Start simple. Venting the containers is absolutely not necessary to have success with tissue culture. It is likely the case that the plants will grow better if the containers exchange air well, but it certainly is not a requirement.

Sterility IS a requirement and having good sterile technique should be the focus for anyone wanting to start tissue culture.
By Apollyon
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Posts:  352
Joined:  Tue May 05, 2020 2:49 am
#362833
Matt wrote:
Jessy2363 wrote:yes it's possible I am overthinking it and becoming obsessional about the need to wanna vent the container.
As a seasoned TC person, I would say that you are. Start simple. Venting the containers is absolutely not necessary to have success with tissue culture. It is likely the case that the plants will grow better if the containers exchange air well, but it certainly is not a requirement.

Sterility IS a requirement and having good sterile technique should be the focus for anyone wanting to start tissue culture.
Do you have any suggestions on where to begin learning the process and how to go about it? Do you have any particular resources you found helpful when you were starting out?
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By Matt
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Posts:  21339
Joined:  Mon Apr 21, 2008 11:28 pm
#363223
Apollyon wrote: Sat Aug 15, 2020 1:38 am
Matt wrote:
Jessy2363 wrote:yes it's possible I am overthinking it and becoming obsessional about the need to wanna vent the container.
As a seasoned TC person, I would say that you are. Start simple. Venting the containers is absolutely not necessary to have success with tissue culture. It is likely the case that the plants will grow better if the containers exchange air well, but it certainly is not a requirement.

Sterility IS a requirement and having good sterile technique should be the focus for anyone wanting to start tissue culture.
Do you have any suggestions on where to begin learning the process and how to go about it? Do you have any particular resources you found helpful when you were starting out?
You know what -- I went looking for a video that was on YouTube and was posted by an acquaintance I had many years ago that was extremely helpful. But it appears to have been taken down.

I also got a lot of help from Carol Stiff at Kitchen Culture Kits way back in the day. I bought her kit and it has good detailed instructions on how to start and comes with everything you need to get started.

If I were to start from scratch and had total amnesia of how to do TC, I'd probably buy one of her kits again:
http://www.kitchenculturekit.com/kits.aspx

Also, the book Plants from Test Tubes was very helpful for me:
https://www.amazon.com/Plants-Test-Tube ... 604692065/

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