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Post by pitcherfreak on Aug 1, 2007 0:32:18 GMT
The best way to do shopping, at your leisure. TC is fun but a bit fiddly. Have tried with other plants and I think looking at the other web sites that contamination is the hardest thing to deal with. You don't know the full name of PPM. Haven't heard of it before here.
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Post by Michael Catalani on Aug 1, 2007 1:13:26 GMT
The best way to do shopping, at your leisure. TC is fun but a bit fiddly. Have tried with other plants and I think looking at the other web sites that contamination is the hardest thing to deal with. You don't know the full name of PPM. Haven't heard of it before here. PPM is Plant Preservative Mixture. Its a biocide that does seem to help reduce contamination, and slow it up when it does occur. This is useful because if a culture is multiplying and then you find a contamination outbreak, you can remove the culture and soak it in this biocide for a while, and then tranfer to fresh medium, thereby saving the culture. It also can be used to soak cultures that are ready to be planted in soil, as these cultures can easily get attacked if there is even the smallest hint of the sugar based medium remaining on them. Here's the website for information on it. www.ppm4plant-tc.com/
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Post by jm82792 on Aug 1, 2007 2:26:04 GMT
Okay I failed tc BUT the contamination was encapulated so it was very hard to tell until the explant turned black yet the culture did not spoil.
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Post by pitcherfreak on Aug 3, 2007 1:11:18 GMT
Thanks JM, will try the site. The explant may not have taken. It may not have been due to contamination. How long did the plant tissue survive? Would love to try with sarr's but it looks like contamination is a big problem with them.
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Post by jm82792 on Aug 3, 2007 4:33:30 GMT
Ask this guy he has done sarr tc sucessfully and he has some growing right now Also the darkness is uninviting until you sign up then it is a white blue color scheme, www.world-of-carnivores.com/cgi-bin/yabb2/YaBB.plHe is very nice and knowledgeable The tissue lived for a few days then turned black so I want to try seeds since the conatmination would be seed by seed.
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agar
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Post by agar on Aug 3, 2007 5:18:56 GMT
Sarrs are actually a breeze from seed compared to tissue, as are most other species (from seed), except for the ones naturally hard to germinate and grow on regular CP soil.
From the sound of it JM, you over-sterilized the tissue, as you had no classic contamination symptoms of contamination - fluffy white (or colored) mold/fungus tufts, cloudy white/tan/pink bacterial growth, or yellow or dark purple phenolic "bleeding" directly adjacent to the tissue. You're tissue just turned dark - and died. Not to worry, I've dumped a few of them in recent weeks.
Contamination should be very easy to detect. Matti's has not only got mold on the explant, but colony's around the dish. This seems to indicate there was likely airborn contamination that occured during transfer, as it doesn't originate from directly from around the explant itself.
Microwave, pressure cooker, or commercial autoclave, isnt so much an issue - this is more of an art than science. Skill and aquired experience is the key. No two people do things the same way, and the results are difficult to interpolate to other situations. MC has good luck with microwaves, but he's 'Dialed in" his process. It works for him. You've just got to find what works best in you're situation;-)
Only 25 B-caps?... I can't remember that far back. I've got several dozens of jars and caps to wash up this weekend;-(
agar
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matti
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Post by matti on Aug 3, 2007 10:46:42 GMT
There was no attempt to tissue culture a plant, it was meerley a test scenario to see what mold would do and if I could grow it.
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agar
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Post by agar on Aug 3, 2007 16:40:45 GMT
Yes, I realize that you were just experimenting, I was just pointing out that the clumps of mold around the leaf, were from airborn contaminants introduced during the transfer, and did not originate from the explant itself. It just looked a bit strange, because I've never seen a case where contamination wasn't directly associated with the explant. Leftovers in the fridge however, are another story;-) Here's a photo of some S. 'Hurricane Creek White' seed cultures I started earlier this spring. A considerable number of seeds germinated, and there's no contamination! Oh, and another advantage of using a pressure cooker or autoclave over a microwave, is that you can sterilize tools (knives, tweezers, forceps) and jars with metal lids;-) JM, maybe try garage sales or EBay. I got my "Presto" 6 quart pressure cooker form Orchard Supply Hardware years ago, and it serves me well. The only drawback, is that it's a tad on the small size. I can only process 7 tall baby food jars (or 14 shorties) at a time. Makes for a busy weekend when you're trying to crank out 50-100 jars;-)
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Post by Michael Catalani on Aug 3, 2007 17:27:41 GMT
There was no attempt to tissue culture a plant, it was meerley a test scenario to see what mold would do and if I could grow it. Unless you live on the moon or inside an active volcano, you are almost guaranteed success in growing contamination on a sugar based medium.
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Post by jm82792 on Aug 4, 2007 5:01:46 GMT
I used PPM in the media so it may have helped I could have over steriliuzed the tissue. I have 30 militers of PPM so I'm going to use that for sterilization of seeds then use bleach since PPM is pricey to give me some condidence. I have 25 caps and only like 15 jars I need more jars ! I want a pressure cooker since you can use metal caps by poking a hole in the (metal)cap and covering with a round band aid. I will be happy when I get some success with tc I'm searching for a pressure cooker since I'm on a tight budget I can't spend $100 on a 26 quart cooker
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Post by pitcherfreak on Aug 6, 2007 0:20:09 GMT
Thanks Agar, don't you just love washing out contaminated flasks/jars. Have you had any success with sarra tissue at all. I know it's hard and seed are far easier but I have a beautiful cultivar that I'd love to have more of, and of coarse it probably won't come true from seed grr.
Thanks JM for that web site have had a look at it is gr8.
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agar
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Post by agar on Aug 6, 2007 4:51:57 GMT
Oh, I haven't *even* started washing jars yet... I topped out at somewhere well over 800-1000 culture jars a month or three ago, and was really squeezed for empties - but, now that I've been deflasking stuff, I now have well over 100 jars to wash up at the moment. And that's after cleaning and preparing 30-40 fresh jars. Current crop of subcultures and transplants include 'fused tooth'. Aren't these little guys cute? Still small, but show the distinct fused tooth characteristic that they are named for;-) Pitcherfreak, no success yet with Sarr tissue, as of yet, but I honestly haven't spent enough time working with them. I've got more experiments lined up though. Had some rhizome tissue I was going to work with earlier this spring, but never got around to it in time. Got a few nice new 'Hummer's Okee Classic' pitchers growing from it now;-) JM, that's another plus about using a pressure cooker. The B-caps vent themselves, so you don't have to mess with covering punched holes in metal lids with Band Aids. But then you couldn't use them in a microwave. I haven't tried using it yet, but NaDCC www.kitchenculturekit.com/nadcc.htmmight be worth a try. Apparently the common pool cleaner tablets available at many discount stores and home improvement centers contain 99% of this chemical. Please refer to the above website, and read and follow the instructions on the bottle if you try out this stuff. Also, if you are a minor, please get approval and assistance from an adult. Seriously guys, be careful with this kind of stuff! agar
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Post by pitcherfreak on Aug 8, 2007 5:50:11 GMT
Thanks Agar. Great photos . Will have to give it a go myself to. Miss having a full blown tissue culture at work like I used to have was handy to say the least. Your cultures look great.
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Post by jm82792 on Aug 9, 2007 3:51:15 GMT
I need to get what I need then I'll go crazy
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