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Post by ischan on May 19, 2008 7:58:02 GMT -5
I have had some success on some of the more straightforward drosera. I wanted to ask if, left alone long enough, do they actually creep onto the agar? Currently they are just filling up all the space from the leaf explant. I don't know if I'm supposed to start cutting them up and placing them into new jars at this point, or wait until they cover the whole surface of the agar. Is there a technique to cutting them up? Or do you just take your knife, sliced into small pieces and dump into new jars? Most of these don't have roots yet. I'm still having trouble with the petiolaris complex. I just can't get them to stay alive inside the media. Within one to two weeks, it turns black in color. Either that, or it looks like a bacteria infection, where white, milky liquid, starts leaking out. I've tried using 1% bleach, 0.5% bleach, and 1ml per 1 L of water. Wash and bleach for five minutes, three baths of clean water after that before transfer into flask. Prior to bleach, use 12 to 20 hour wash with water, change at least three times. My media is 25% MS. is my media still too strong? I had trouble getting some of the Sun dew to grow using 50% media, until I was corrected by some of the folks on Dr. Carol's list serve. Finally, I have some tillandsia seed growing in TC. Anybody tried this before? I havent a clue what to do with it now!
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Post by lloyd on May 19, 2008 12:27:56 GMT -5
I'm impressed! All my explants have oxidized, never grown or died from contamination. I've only had some success from seeds.
You don't have to wait for roots to divide plants in TC. From what I've read when they have roots they stop dividing and can be taken out of TC and hardened.
I had some D. derbyensis growing in agar from seed. I left them in too long and they died of shock when I took them out. Also I tried some leaf pulls and they all oxidized and died.
Post your techniques, sterilization, media ingredients, etc; I'd like to see what you use.
1/4 to 1/2 MS sounds good by the way.
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Post by vraev on May 19, 2008 13:28:37 GMT -5
fantastic man! u can start up your own business with those skills. Well done.
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Post by ischan on May 21, 2008 18:17:46 GMT -5
Nothing special really, these are the tough weedy CPs. If I can get it working on the difficult plants I'll be happy.
1% bleach, 0.5% bleach, and 1ml per 1 L of water. Wash and bleach for five minutes, three baths of clean water after that before transfer into flask. Prior to bleach, use 12 to 20 hour wash with water, change at least three times.
Thats all I'm doing, and MS media
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Post by lloyd on May 21, 2008 19:13:19 GMT -5
By 1% bleach do you mean 1 cc of household bleach to 99 cc of water? Also do you mean you use three 5 minute washes of each of the 3 bleach concentrations? Thanks.
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Post by ischan on May 24, 2008 8:14:49 GMT -5
I use 500 ml water, and add 5ml bleach or 2.5ml so its not quite 1% but its there somewhere. my measuring bowl for 500ml isnt quite that accurate either. its a baking kitchen bowl
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Post by lloyd on May 24, 2008 13:50:17 GMT -5
I was usiing 10 cc bleach to 90 cc water for a 10% solution. I'll have to try the more dilute concentrations.
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Post by ischan on May 25, 2008 17:27:24 GMT -5
I never had explat live at >1% bleach. Even 1% is a lot. But at such weak bleach your contamination is also high. It's just that way...
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Post by lloyd on May 25, 2008 19:10:57 GMT -5
Do you use PPM?
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Post by panduka2007 on Dec 18, 2008 20:02:29 GMT -5
hi all please let me know from wher i can buy tissue culture media and aga or phytagel and hormones from TORONTO, TNX MAHEN
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Post by hackerberry on Dec 18, 2008 22:15:16 GMT -5
Hello Mahen,
We order online for the tc stuff. Phytotechlab. com.
Cheers,
hb
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Post by lloyd on Dec 19, 2008 8:16:46 GMT -5
Hi Mahen, I pm'ed you. If you come in to Toronto, I can give you some stuff to start with. I could mail it but the post office charges are annoying.
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