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Post by flytrapcare.com on Nov 8, 2009 23:43:17 GMT -5
Ok, so my first successful explant was a Cupped Trap leaf. Here's a photo of what I started with back in March: Here's the original thread I started when I though it was making callus: www.flytrapcare.com/phpBB3/viewtopic.php?f=28&t=763Well, I'm absolutely stunned what I was able to turn that one leaf into over the last 6 months. Check it out: That's only about half of the jars I have right now and I've also sent some to friends. I also deflasked quite a few back in September. Some of them I've sent to friends or FlytrapCare Store customers. Recently I deflasked a bunch more. In all, what you see here is less than 1/3 of all of the Cupped Trap plants grown from that one leaf. It was a very long day of potting to say the least, but tissue culture definitely works!
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Post by hackerberry on Nov 9, 2009 7:57:08 GMT -5
Well done Matt!
hb
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Post by Devon on Nov 9, 2009 8:11:46 GMT -5
WOW! that was all from one leaf!? That's awsome!! I would definetly like to try TC some day... sme day. Very nice thanks for sharing
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Post by lenynero on Nov 9, 2009 10:48:39 GMT -5
Hi,
Well DONE!!!
I noticed that you changed media to one that contains charcoal. Is it your own ingredients or from a mix?
Thanks,
Leny
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Post by flytrapcare.com on Nov 9, 2009 17:58:46 GMT -5
Thanks hb, Devon and Leny!
Leny, I added 1g/L of activated charcoal (I just purchased it at a health food store in capsules) to the grow media. The charcoal is supposed to help absorb any of the residual plant growth regulators.
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Post by lloyd on Nov 9, 2009 20:18:14 GMT -5
That is incredible. I've never had any success with explants. Could you go over your sterilization procedures, agar recipes and any helpful hints?
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Post by flytrapcare.com on Nov 11, 2009 1:53:27 GMT -5
Could you go over your sterilization procedures, agar recipes and any helpful hints? I use the same basic sterilization procedure for all seeds and explants. I just vary the time depending on the size of tissue or seeds. It's basically this: 1) Wash the explant well with soap. Antibacterial soap is best. Sometimes I let it soak a bit depending on the "robustness" of the tissue. 2) 15 to 30 second dip in alcohol (I might skip this if the tissue is sensitive or small) 3) 2 to 5 minutes in H2O2 4) Quick dip in sterile water to get the H2O2 off. 5) 6 to 15 minute soak in 10% bleach with 2 to 3 drops of Tween 20 per 100ml. 6) Two good rinses in sterile water for 3 to 5 minutes. For my media, I always use 50 MS with full strength MS vitamins, 1 g/L of PPM (for explants and seeds -- not needed for sterile cultures), 30 g/L sucrose, pH = 5.6. That's it! As for the agar, I add it to the baby food jars individually. I put a smidgen (from a smidgen spoon measuring set) per 25ml and a dash for 45ml. Hope that helps!
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Post by lloyd on Nov 11, 2009 9:32:36 GMT -5
That's pretty well what I have done, except I added hormones to my explants which I suspect were toxic to the sensitive CP's. Thanks for the notes and congratulations on your success.
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Post by flytrapcare.com on Nov 11, 2009 10:24:17 GMT -5
Thanks lloyd! Yeah, I've definitely noticed a correlation between PGRs and explants turning black. I stay away from them entirely until I'm ready to propagate tissue. For establishing explants, they're not needed.
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Post by auth100488 on May 13, 2010 17:27:50 GMT -5
Impressive can i ask something?, since your jars are kind of big, you are probably not using a pressure cooker, what have you been using ? thx
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