Tissue Culture FAQ

Discuss all micropropagation related topics here.
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Matt
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Re: Tissue Culture FAQ

by Matt » Tue Jan 05, 2010 7:16 pm

Xeno wrote:Very nice Matt since it wasnt that long since you did start from what I can recall.

Yeah, I only started a year ago in November. At least, that was my first attempt. I didn't really start in earnest until around March, when there was more tissue to work with. This summer was a tough one for me to get too much going with TC because my wife and I made a very long move. This summer will be much better and I'm optimistic that I'll be able to get most of the clones from my collection started in TC.
Xeno wrote:Its then possible to have one in a jarr for a long time for later use and then I could use that one for explant since its already clean ?

Yes, once you get a clone started in TC, it's very easy to keep it going indefinitely. Some people say that you should restart them from time to time because you can get genetic mutations if you've divided them too much. But then I've heard other people say they've propagated the same clone for 15 years without any issues.

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Re: Tissue Culture FAQ

by Noddy » Fri Jan 15, 2010 11:00 pm

Hi can you advise what vinger to use as there's a few types... To adjust the PH level?

Thanks Noddy

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Re: Tissue Culture FAQ

by renesis » Fri Jan 15, 2010 11:18 pm

Noddy wrote:Hi can you advise what vinger to use as there's a few types... To adjust the PH level?

Thanks Noddy


Normal white vinegar, acetic acid.

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Re: Tissue Culture FAQ

by goldslinger » Sun Jan 17, 2010 5:49 am

Is KOH and Muriatic (HCL) acid often used because it is more stable over time in the media? The PH won't change as they are better buffers? Someone mentioned they were more reliable, albeit MUCH more dangerous. Is Vinegar and Baking Soda less inert as well?

I can get Muriatic acid at the hardware store. I think it is 36%, don't know for sure. If I get the KOH, I will get the solution if it has a good shelf life.

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Matt
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Re: Tissue Culture FAQ

by Matt » Sun Jan 17, 2010 6:09 am

goldslinger wrote:Is KOH and Muriatic (HCL) acid often used because it is more stable over time in the media?

I've not heard this, but I'd like to know if it's true. I have some KOH that I use as a solvent when dissolving PGRs, but I always use baking soda for raising the pH of media. Is it better to use KOH?

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Re: Tissue Culture FAQ

by renesis » Sun Jan 17, 2010 11:59 pm

goldslinger wrote:Is KOH and Muriatic (HCL) acid often used because it is more stable over time in the media? The PH won't change as they are better buffers? Someone mentioned they were more reliable, albeit MUCH more dangerous. Is Vinegar and Baking Soda less inert as well?

I can get Muriatic acid at the hardware store. I think it is 36%, don't know for sure. If I get the KOH, I will get the solution if it has a good shelf life.

Gary


I'm speculating people say its more reliable because the strength will be the same every time, whereas something like baking soda may contain other impurities batch to batch. But, as for being more stable in media, I doubt it. Both baking soda and KOH would break down into their ionic forms and contribute to the overall basicity the same way. Adding more potassium (KOH) instead of sodium (baking soda) on the other hand might make a positive difference though. :?:

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Re: Tissue Culture FAQ

by mfdes » Tue Mar 02, 2010 1:35 am

Hi guys and gals, a couple of observations:

I have used gelcarin in the past. Supposedly less likely to induce hyperhydricity. Dunno, I could never tell.
However I like that it's optically clear. One thing to note is that it is more brittle when set than agar, so shipping cultures is a more delicate affair.

Secondly, when it comes to pressure cookers, pay attention to the rated PSI. You might have to look in the fine print. I learned the hard way that what is considered standard by one retailer (15 PSI in most of the U.S., where home canning is popular), ain't necessarily what another retailer considers standard. Here in Australia 7 PSI pressure cookers are regularly sold WITH NO MENTION OF THEIR OPERATING PRESSURE in the retail information package. I bought one and only found out when I rang up to enquire after food was taking longer to cook than I had expected. Be warned. Needless to say I was stuck with a $200 machine that I could not return because (legally) there was nothing wrong with it and it had already been used.

Awesome site by the way!

M

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Matt
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Re: Tissue Culture FAQ

by Matt » Tue Mar 02, 2010 6:14 pm

Thanks for the feedback mfdes. Very helpful information.

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Re: Tissue Culture FAQ

by robert128 » Sun May 23, 2010 1:44 am

Hi,great post
I need some help with a unit of measurment.
I want to try a new TC media formula i have found on the net but the measurments use the prefix uM as a unit of volume but googling this unit has come to a dead end.
I think it means micrometer but this is a unit of length not volume or mass.
so does 1 uM = 1 microgram ?

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Matt
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Re: Tissue Culture FAQ

by Matt » Sun May 23, 2010 2:09 am

uM is a micromole. It's determined by the molecular weight of the compound you're dealing with. If you want to convert micromoles to milligrams, see here:
http://wiki.answers.com/Q/How_do_you_co ... milligrams

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Re: Tissue Culture FAQ

by jht-union » Mon Jun 14, 2010 3:46 pm

for sterilizing seeds, can it be distilled water, when rinsing the seeds?


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Matt
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Re: Tissue Culture FAQ

by Matt » Mon Jun 14, 2010 4:55 pm

Any rinses done after the sterilization process should be done in sterile water. Before the sterilization process, it doesn't matter too much what kind of water you use, but the cleaner the water is, the better.

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Re: Tissue Culture FAQ

by Doomsday » Tue Jun 22, 2010 4:59 am

Do i need to put ppm into the water before i microwave it to make it sterile, or could i just bring it to a boil for a minute or two
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Matt
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Re: Tissue Culture FAQ

by Matt » Tue Jun 22, 2010 6:07 am

PPM isn't needed to sterilize water. Just bring it to a boil. The amount of time depends on how much water is in the jar. I'm not familiar with times when using a microwave. But for a pressure cooker, I usually sterilize my 150ml water flasks for 30 minutes.

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Re: Tissue Culture FAQ

by Hayden » Tue Aug 31, 2010 4:12 pm

Hi, im looking to start tissue culture next year if possible! Im not looking for anything massive, only something I can fit about 10-20 jars in at the most! Like all the media and accessories sounds fine, and fairly cheap. But im wondering where and how I can get a decent tissue culture 'zone'.

Thankyou :)

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