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OfflineMyOwnReality
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Petri dishes and agar
    #7936281 - 01/26/08 05:34 PM (16 years, 6 days ago)

So I have a question, what kind and size of petri dishes are best for doing tissue cultures? I.E. Glass or plastic, thickness etc. What agar formulations are best for culturing cubensis(right now I'm using malt/dextrose) What about vented plates?

Also any links to articles on tissue culturing and strain isolation would be appreciated(I've found some but I'd like other people's opinions.


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Invisibledumbfounded1600
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Re: Petri dishes and agar [Re: MyOwnReality]
    #7936486 - 01/26/08 06:16 PM (16 years, 6 days ago)

I like the plastic because there ligher and not as slippery as the glass can be when wet


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Invisiblefastfred
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Re: Petri dishes and agar [Re: dumbfounded1600]
    #7942878 - 01/27/08 09:36 PM (16 years, 5 days ago)

There is a thread on tissue culture here in advanced. It should still be on the first page of threads.

> what kind and size of petri dishes are best for doing tissue cultures?

Standard dishes are 100mm x 10, 15, or 20mm. Stackable ones are easier to work with. If you get the shallower dishes it's easier to pour the media up to the top so that you can look at them under the scope through the dish.

> Glass or plastic?

They each have their uses. Glass are reusable and can be autoclaved with media in them. That makes them worthwhile IMHO. Plastic are useful because you can get different sizes and triple sectioned ones. RR likes the sectioned ones.

Plastic are about 15-20 cents each, glass are around 85 cents each. So if you use your glass ones at least 4 times they will come out cheaper.

Cynmar is the cheapest place for science supplies that I've found.

GLASS PETRI DISH, 100x22mm,10/PK: $8.65
http://www.cynmar.com/item_detail.aspx?ItemCode=13220698

> What agar formulations are best for culturing cubensis

MEA and PDA are the best for edibles IME. They're both easy to make and dirt cheap. Some people add yeast extract and think it helps a little. I don't use it except for special circumstances. Check out my media cookbook in the agar section here.

> What about vented plates?

Never used them. Don't see why you would. Most people go a step in the other direction and parafilm their dishes. It makes them easier to handle/examine, keeps contams out, and keeps the media from drying out.


-FF


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Offlinebw86
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Re: Petri dishes and agar [Re: fastfred]
    #7965480 - 02/01/08 04:03 PM (16 years, 10 hours ago)

1.5 inch plastic dishes


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InvisibleArp
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Re: Petri dishes and agar [Re: bw86]
    #7965760 - 02/01/08 05:05 PM (16 years, 9 hours ago)

i use glass. easy to sterilize, and will last a lifetime if treated properly


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InvisibleYidakiMan
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Re: Petri dishes and agar [Re: fastfred]
    #7967434 - 02/02/08 01:22 AM (16 years, 1 hour ago)

Quote:

fastfred said:
Cynmar is the cheapest place for science supplies that I've found.






I was a big fan of cynmar until they refused my last order because my credit card statement is sent to my parents house. I was a repeat customer. I ordered from them 4 or 5 times and they refused to do business with me anymore.


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Invisiblear393
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Re: Petri dishes and agar [Re: YidakiMan]
    #8059694 - 02/23/08 09:50 AM (15 years, 11 months ago)

cenmed is where I get them from. I use plastic.

And after a few years away from the hobby, I just started some plates a few weeks ago and just inoculated some popcorn....its good to be back.


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Offlinesolumvita
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Re: Petri dishes and agar [Re: ar393]
    #8077219 - 02/27/08 01:24 PM (15 years, 10 months ago)

I find that using a 1 % PDA is more effective for tissue work as the mycelium does not get as thick and it is easier to do hyphal tip isolations from the sparse mycelium. but once you have a clean strain then full strength PDA or MEA is best for vigorous growth.


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OfflineMycoAu
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Re: Petri dishes and agar [Re: solumvita]
    #8077360 - 02/27/08 02:12 PM (15 years, 10 months ago)

I am about to blatantly show my ignorance, so tread lightly.  :blush: *ignorance exposed*

Why hyphal tip isolation over some other method?  I presume this means trying to select one (or as few as possible) strands of the mycelium, from the newest growth regions in a sector that is currently displaying favorable characteristics.  Is it just to narrow down the isolation time because there are fewer possible genetic pools to deal with if you can isolate a single strand?  :blush:


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Offlinesolumvita
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Re: Petri dishes and agar [Re: MycoAu]
    #8081239 - 02/28/08 12:01 PM (15 years, 10 months ago)

stupidity is not asking :smile:, i just used hyphal tip isolation, to illustrate the use of half strength PDA.  I use it more when trying to recover a contaminated strain, as you can then literally just get the tip of the strand and be fairly sure that it is free of contaminants.  of course if there are bacteria involved you would also need to use an antibiotic in the agar to remove the bacterial contam. 
You actually need just the tip of the mycelia, the whole strand would be too much.  hope that makes sense.:cool:


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InvisibleInfiniteOhms
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Re: Petri dishes and agar [Re: fastfred]
    #8081484 - 02/28/08 01:23 PM (15 years, 10 months ago)

Quote:

fastfred said:
Cynmar is the cheapest place for science supplies that I've found.




Thanks for that, i ordered 20 glass dishes ... and saved some $ over the other place i was going to order from.


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OfflineMycoAu
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Re: Petri dishes and agar [Re: InfiniteOhms]
    #8081607 - 02/28/08 01:55 PM (15 years, 10 months ago)

Ok, so hyphal isolation seems like a good idea and all, but... How is it that you're recovering just one tip? Just eyeballing it? Better equipment? Magnification of some flavor? Under a microscope in the hood? Just wondering...


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Invisiblefastfred
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Re: Petri dishes and agar [Re: MycoAu]
    #8081633 - 02/28/08 02:04 PM (15 years, 10 months ago)

I've found pretty decent results using a 1 mL pipet tip under up to 100X.


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OfflineMycoAu
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Re: Petri dishes and agar [Re: fastfred]
    #8081681 - 02/28/08 02:21 PM (15 years, 10 months ago)

Ok, so the pipette tip isn't available to me. Do you see any problems using a 25 to 27G needle?


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Invisiblefastfred
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Re: Petri dishes and agar [Re: MycoAu]
    #8084847 - 02/29/08 02:54 AM (15 years, 10 months ago)

Try an eyedropper perhaps. It's handy to know that it's the second line that is the bottom and actually watch your target get pushed up into it. I've actually done some at 400X and it's not that easy unless you can really see what you're doing.

Those pipette tips are generated by the thousands by most biology labs. They are disposable, but actually made to be autoclavable, and usually get autoclaved prior to use and then thrown away afterwards. Glass pipettes also would work well I imagine.

See:


http://www.cynmar.com/item_detail.aspx?ItemCode=13534784
and

http://www.cynmar.com/item_detail.aspx?ItemCode=13224551


-FF


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OfflineMycoAu
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Re: Petri dishes and agar [Re: fastfred]
    #8085150 - 02/29/08 07:45 AM (15 years, 10 months ago)

It's not a problem getting those companies (Fisher/Aldrich, etc.) to mail to your house? Last time i checked, they wanted a business mailing addy. I could probably use the one at work, but my boss might also get really pissed- who knows?

I'm thinking the glass ones are more my flavor despite the larger pore size in the tip. Glass and metal are my favorite materials because I know that they are chemically and thermally resistant with a little care.

Any other good tips for a budding "mycologist"? Thanks to everybody here offering their help and especially FF!


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Offlinesolumvita
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Re: Petri dishes and agar [Re: MycoAu]
    #8086068 - 02/29/08 12:58 PM (15 years, 10 months ago)

I use a dissecting microscope up to 320X on a laminar flow bench and then a really steady hand and some good lighting to see the mycelia and a sharp needle.  But i think i might try the pipette tip, thanks fastfred :thumbup:


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Invisiblefastfred
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Re: Petri dishes and agar [Re: solumvita]
    #8087714 - 02/29/08 08:09 PM (15 years, 10 months ago)

> It's not a problem getting those companies (Fisher/Aldrich, etc.) to mail to your house?

Unless you want chemicals cynmar will ship to you just fine, easy transaction. If you want chemicals from them they want some sort of business info, again not a problem if you have a business tax ID # or some sort of paperwork.

It's no hassle for any of the normal sort of lab supplies you might need. Inoculating loupes, test tubes, media, beakers, parafilm, petri dishes, etc..


-FF


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OfflineMycoAu
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Re: Petri dishes and agar [Re: fastfred]
    #8091447 - 03/01/08 08:12 PM (15 years, 10 months ago)

Good to know. A few years back, I wanted some chemicals and was going to ship to my house, but that's when I heard about the business address requirement. I was unaware of general lab supplies being available without one. This is great news for a few of the other materials, etc. that are hard to find a DIY replacement.


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