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Post by pyrgoscper on Mar 24, 2007 3:52:08 GMT -5
Hello, I am totallty new to the world of TC, (its incredible if you ask me!) I have been reading and reading etc, but i was wondering if someone can help me out? I know seeds need to be sterilized and put in media etc etc, I bought a kit from kitchenculturekits WICH I highly recommend!! ANd i am lost within the universe of cp tc! If possible could someone tellme an exaple from start to finish on how to tc a P.esseriana from a leaf section in tc? Any help will be greatly appreciated, thankyou! I do have test tubes, media and a laminar flow hood but where to begin åõ÷áñéóôþ Thanks!
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Post by sdeering on Mar 24, 2007 8:31:45 GMT -5
There is a plethora of info on the net if you look. Sterilize the leaf in a mixture of 10% household bleach to water for 10 to 15 min. Under sterile conditions rinse it in sterilized water 3 times and place it on your favorite sterilized medium. See wasn't so hard. For some reason I can't seem to get the leaf sterile. It usually has contaminants inside that end up taking over, so don't be surprised if you get a nice growth of not Pinguicula but mold, bacteria and fungus. Good luck.
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Post by pyrgoscper on Mar 24, 2007 12:41:39 GMT -5
thanks i will give it a try! how long do i wash them off in sterile water for ? like 5 mins at a time x3?
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Post by sdeering on Mar 24, 2007 13:39:42 GMT -5
I usually rinse for a minute or so 3x.
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Post by lloyd on Mar 24, 2007 19:57:53 GMT -5
I've had some problems with systemic contamination and also phenolic bleeding. I'm trying decontamination using PPM 4% in full strength MS in distilled water for 4 hours and then putting the material right into the medium.
If you go through the TC section here, you'll find lots of good advice and links.
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Post by pyrgoscper on Mar 25, 2007 3:48:36 GMT -5
thankyou very kindly for the help, i will give it a try (gotta give this darn laminar flow hood a run for the money so to speak lol
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Post by pyrgoscper on Mar 28, 2007 0:33:40 GMT -5
Well ive went out and bought all the misc supplies and a pressure cooker for 6$ at Valu Village of all places and i tc'ed or flasked i guess i should say a bunch of p esseriana and b-52 flytraps so hopefully all goes well and i will post the results and hopefully a picture soon!! thx for the help!
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Post by lloyd on Mar 28, 2007 11:27:17 GMT -5
Did you make or buy your laminar flow hood? Could you post a picture of it if you made it? You're probably the only one here who has one. Good luck with your TC. What ingredients did you use for your medium?
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Post by sdeering on Mar 28, 2007 14:28:42 GMT -5
I made one Lloid if your interested in the construction I can post a pick and some imfo.
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Post by lloyd on Mar 28, 2007 15:46:43 GMT -5
I didn't know anyone here had one. I'd like to see your pictures and any plans you have.
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Post by pyrgoscper on Mar 28, 2007 17:26:09 GMT -5
Did you make or buy your laminar flow hood? Could you post a picture of it if you made it? You're probably the only one here who has one. Good luck with your TC. What ingredients did you use for your medium? I bought my laminar floow hood off a friend for $200 i thought hey if all goes well with my tc experiments. and the media im using is 1/2 strength ms with sucrose, BAP , and PPM, IBA and i use 1/4 of a teaspoon agar per flask. ;D
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Post by pyrgoscper on Mar 28, 2007 17:27:22 GMT -5
oh and i will put some pics up asap thx
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