Thread: Tissue Culture
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Old 08-31-2012, 09:42 PM   #79 (permalink)
dinhml
 
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Default Re: Tissue Culture

Hi guys, Here's my 1 day reading


it's not that difficult as you think it is. try it. it should be a fun and also a challenge. different authors have different formulas, but it all boils down to.


find all necessary utilities, P.H. meter, pressure cooker, micropipettes, 250ml flash, electronic scales, forceps


steriles all of these in pressure cookers @ 250F, or with strong ethanol or isopropyl alcohol

ingredients:

1 L of de-ionized water

4.3 gram of MS (Murashige and Skoog salts)

5ml of BAP, sometimes called BATH for nutrients purposes(studies show 5ml of BAP is the minimum to get the maximum of plantlets out of the test tubes or whatever containers)

1ml PPM(plant preserve material) for antimicrobial purpose (i think imma experimenting skipping this PPM step) - BUY HERE


AGAR, which is jelly that comes in patri dishes in most labs, which i think the powder form can be obtain thru the supermarkets for making jelly, which then u can boil with water and make AGAR out of it

Add Sugar and coconut milk for experimenting, since all formulas are self-made by a guy in his lab, so can we too . i suspect u add sugar in this step if u gonna mix the powder agar with water by yourself. Coconut milk acts as a growth promotor so add it if u have any.
add all the stuffs above together in a sterile flash/container and measure the P.H., try to have it 5.8, if PH too high, add vinegar, if too low, add antacid tablets. PH is crucial, the exacter to 5.8, the better


then boil in pressure cooker or microwave it until everything boil

let the mixture cool

pour in jars (where explant gonna go)

_____________________________
FOR the Corm/or Explant

trim it to 1 inch if it is from a flower, bigger size for corms

dipped it in bleach solution with 2 drops of surfactant (dish soap) for 20 minutes, then wash 3 times with deionized water

remove again the outter tissues since it got damaged by the bleach, now u have a perfect living sterile explant

when trimming, remember to use sharp razors or knives to avoid dulling the tissue

transfer this explant to the prepared jars contained the mixture, close the lid or by parafilm if test tubes are used.

let it sit under flourescent light for 16 hours daily at room temperature 75-85F


Most important of all, KEEP EVERYTHING CLEAN FROM BEGINNING TO END

AND REMEMBER: Not all formulas are exact, try making your variation, u dont have to be exact


and lastly

report your success with formulas back on this page

have fun growing ALL

USEFUL LINKS

: watch all of his series

this has 3 parts so find it

learn aseptic techniques from youtube

read tissue culture for home gardener there ain't error, wait 1 minutes before page load

and read anything that is on tissue culture on bananas.org


and read Ian Mcguire

LASTLY: any inputs to make the process shorten is always appreciated

Last edited by dinhml : 08-31-2012 at 10:25 PM.
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