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Post by StickyIcky on Apr 9, 2015 14:21:26 GMT -5
I've recently been promoted, and am now manager of my own little facility. So I now have my own autoclave, biosafety cabinets, hot bead sterilizers, flasks/plates and what not. I've had access to these for years at my old facility, but had a direct supervisor to answer to. Now that I'm my own boss (at my facility at least), I'm thinking of getting the other necessities together to do some TC work during my breaks/after hours. I guess I mostly just need the medium at this point. Kinda excited! Sterile work is something I do daily, so I figure I should be able to manage. I think my biggest concern is tissue sterilization (Without killing the tissue). Start with seed?
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Post by lloyd on Apr 9, 2015 15:59:19 GMT -5
Congratulations. I'm jealous! If you have to retire any equipment let me know if I could buy something.
Seed is definitely easier to sterilize without killing with some provisos. VFT seed has to be hulled (gently) first. Otherwise mould inside the shell will infect the medium AFTER germination which is really annoying.
There are lots of protocols, 10% bleach for 10 minutes (with Tween 20) is reasonable.
Explants are also worth trying although in my hands at least, much harder. I have found that CP explants are super sensitive to PGR's and you should use 1/10 the usual concentrations. You can use the bleech for explants too but with more care especially with thin or delicate tissue.
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Post by amanitovirosa on Apr 9, 2015 16:35:03 GMT -5
...Congrats on the new gig, I know nothing about tissue culture.
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Post by shoggoths on Apr 9, 2015 18:49:53 GMT -5
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Post by StickyIcky on Apr 9, 2015 19:29:03 GMT -5
Awesome! thanks guys! I'll definately read up a ton more in the meantime. Still gotta get my bearings around the place, and then once I'm a little more in the comfort zone, I'm going for it.
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Post by pinhead on Apr 9, 2015 20:00:10 GMT -5
I have done some research and have compiled a file with protocols from the web and ones that have worked for me.
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Post by pinhead on Apr 9, 2015 20:12:26 GMT -5
I should mention the file is a Word file.
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Post by roraima on Apr 10, 2015 2:08:07 GMT -5
Congrats!
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Post by StickyIcky on Apr 10, 2015 11:19:16 GMT -5
Thanks Guys! And thanks Guy
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Post by Devon on Apr 10, 2015 12:28:43 GMT -5
Congrats! I hope you post pictures of your TC endeavors when you get it going.
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Post by sokkos on Apr 10, 2015 15:24:19 GMT -5
Congrats! Take lots of pictures of your progress!
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Post by H2O on Apr 10, 2015 15:39:26 GMT -5
Always great to have another TC man on our forum!
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Post by vraev on Apr 11, 2015 0:38:16 GMT -5
Fantastic man. Looking forward to seeing some of your exploits with TC. I agree...start with some seed. I have gotten decent success (not always...would say 50/50 ) with eBay nep seed. check it out. I have access to all lab related stuff at school. Unfortunately it is just for school work. I don't think they will appreciate me growing/culturing nepenthes/cephs/vfts in BSL LVL 2 / 2+ labs. We pretty much have to stick to cell lines or primary tissues.
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Post by LucST on Apr 11, 2015 1:00:32 GMT -5
I dunno vraev a lot of the profs at VIU are pretty cool and are willing to entertain cool projects. That being said I don't know what the biology department is like, and the botany portion of it will be nil to none.
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Post by vraev on Apr 11, 2015 1:08:36 GMT -5
I dunno vraev a lot of the profs at VIU are pretty cool and are willing to entertain cool projects. That being said I don't know what the biology department is like, and the botany portion of it will be nil to none. Perhaps in the bio dept it would fly...but not in the health sciences dept. Especially, if there ends up being any contamination issues, you can see the blame directly coming towards me. Especially because all our protocols need to be approved by the biosafety office.
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