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hamloaf
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Registered: 12/23/09
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Re: Potato Dextrose Agar (PDA)! [Re: Javadog]
#12971727 - 07/29/10 12:44 PM (13 years, 7 months ago) |
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Cool. 750 milliliters is quite a lot of agar. The idea in the amount of agar I prepare in this tek is to prepare enough so if in your first few attempts at pouring agar, if you over pour your dishes, you won't run out of agar. So go with the amounts in the tek until you get your agar pouring technique down. Using the amounts in my tek, you will have agar left over. HL
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FractalXplora
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Registered: 02/11/06
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Re: Potato Dextrose Agar (PDA)! [Re: hamloaf]
#13025050 - 08/09/10 03:00 AM (13 years, 7 months ago) |
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ham, im just about to try this tek, gonna do half measuers as i dont need 25 plates full.
Ive got no distilled water, was gonna use tap water thats been boiled, can i mix it up while the tap waters still hot\?
cheers.
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Mephistophelian
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Registered: 08/14/08
Posts: 2,527
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Tap water is more then fine for use with agar. The biggest point of using distilled water is for spore syringes so that the spores don't germinate inside the syringe.
So don't worry about a thing. Just do it up.
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Base Icks
Registered: 03/19/10
Posts: 6,191
Loc: Shroomshire
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Just make sure you got enough for your 20 plates
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FractalXplora
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Registered: 02/11/06
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knocked up a half recipe, set like a goodun.nice tek Hammy
Think I let it cool too much (1 hr 40 mins) was slighty gloopy / lumpy when it came out to pour, still no condensation issues like my last run, glad the tap waters ok, all done in a glove box so fingers crossed.Gonna knock em straight up then parafilmed so I can breath easy again.
Next time I think I'll feel the correct pouring temp, I'll give it like an hour 20 mins should be perfect. Still got 11 dishes to practice with for my next attempt.
Have to say its a pure joy knocking up your own PDA, cracking fun!
Doing agar petris takes practice yes, i will get it down, i will get it down, i will get it down
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Base Icks
Registered: 03/19/10
Posts: 6,191
Loc: Shroomshire
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I think most people pour all the plates from a sleeve at once, then put then all back in the sleeve to keep them sterile ish. I read that they dont keep long unpoured. at least when they are poured you can see if they are contaminated.
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Mephistophelian
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Re: Potato Dextrose Agar (PDA)! [Re: Base Icks]
#13025685 - 08/09/10 09:25 AM (13 years, 7 months ago) |
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I personally wrap the column in Glad-Wrap after its poured to add an extra layer of protection while they are in waiting. If you have spares that are un-poured you can do the same thing. Make sure your thorough and always use your sleeve after wrapping the column.
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hamloaf
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Re: Potato Dextrose Agar (PDA)! [Re: hamloaf] 1
#13025802 - 08/09/10 10:04 AM (13 years, 7 months ago) |
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Fuckin a kids. I'm a way for one second and...... Good work, Mephistophelian on fielding these questions while I was away. Way to be! Tap water's fine for agar. Distilled water is just a personal preference. I like to start with a clean slate. Distilled water's true purpose is for making spore solution. Because disstilled water has no nutrient content due to the distilling process, spores will have nothing to feed upon and take linger to germinate for purposes of long term spore solution, as stated above. You could even use pond water. Once through the PC it all becomes sterilized just the same. Ky jelly is good to add to spore solution before the PC at 2 drops per 100 liters of water as well. It keeps the spores form clumping together over longer periods of storing time. Well done, FractalXplora
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Think I let it cool too much (1 hr 40 mins) was slighty gloopy / lumpy when it came out to pour,
Pour your agar when it has cooled to the point where the container is still hot but can be handled and the agar is still liquefied. It's a balance that you can only achieve through practice. Well done, FractalXplora HL
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FractalXplora
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Registered: 02/11/06
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Re: Potato Dextrose Agar (PDA)! [Re: hamloaf]
#13025870 - 08/09/10 10:18 AM (13 years, 7 months ago) |
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sure thing Hamloaf, its only my second attempt at pouring proper plates, fooked it up last year good and proper, got the fear and turned to cooking ready mix agar in jars to do my agar experiments.
Thought about doing all 20 but had a feeling something may go slighty wrong aka gloopy agar.
Didnt want to put em back in the sleeve un-parafilmed as it would maybe mees my 11 clean plates up. So i just left in the glove box for a hour under a new cut down white bin bag (should be sterile on the inside i imagine/hope!
Have i got this right, if you do put straight back into sleeve for storage you don't parafilm them until you got em back out and you've transferred something into dish yes?
Clean ones should be ok for a week or so? wrapped an elastic band round sleeve real tight after opening.
Knocked up 2 ape, 2 shiitake, 2 mex A, and 2 king oysters.
We shall see how clean I was.
lol, one of the MEx A i managed to get the chunk stuck to the dishes lid after parafiming.Urgh! Clever myc will find a way down Im sure.
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Mephistophelian
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Quote:
Mephistophelian said: I personally wrap the column in Glad-Wrap after its poured to add an extra layer of protection while they are in waiting. If you have spares that are un-poured you can do the same thing. Make sure your thorough and always use your sleeve after wrapping the column.
No need to spend a wad of your parafilm wrapping them individually. Save your film for when you go to use them. Just use Glad-Wrap after pouring (while hot if you like) then put your sleeve over the stack then rubber band/clip/seal. Wrap the whole column solid then just cut the dishes out as you need them. That way they stay 'individually wrapped' until you need them.
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hamloaf
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Quote:
if you do put straight back into sleeve for storage you don't parafilm them until you got em back out and you've transferred something into dish yes?
I place the tower of petris back in the sleeve unparfilmed then put the parafilm on after inoculation.
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Clean ones should be ok for a week or so? wrapped an elastic band round sleeve real tight after opening.
Not sure what you mean by "clean ones", wrap your tower of petri dishes in saran wrap for long term storage. HL
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FractalXplora
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Registered: 02/11/06
Posts: 2,494
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Quote:
No need to spend a wad of your parafilm wrapping them individually. Save your film for when you go to use them. Just use Glad-Wrap after pouring (while hot if you like) then put your sleeve over the stack then rubber band/clip/seal. Wrap the whole column solid then just cut the dishes out as you need them. That way they stay 'individually wrapped' until you need them.
Sounds like a plan
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The White Rabbit
Registered: 03/30/10
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I tried doing the saran wrap thing but just destroyed a bunch of dishes when I knocked them over. Now I just leave them in my Glovebox with Alcohol soaked paper towers covering the arm holes. And I spray the shit out of my glove box with Lysol before and after pouring my dishes. So far no contams even up to a week sitting in my glovebox. Only seem to get contams after I transfer or spore swipe.
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The White Rabbit
Registered: 03/30/10
Posts: 257
Last seen: 13 years, 2 months
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I also love parafilm. I got the 2" x 250 and then I cut them into strips. I cut sheets of three squares then cut then in half so they are now 1" X 6". This covers the edge of the petri and yet I can still see the whole dish.
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Mephistophelian
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Registered: 08/14/08
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Ya, Parafilm is just amazing...I'm a recent convert as well.
Got a deal on some 4" x 250'. Just cut a little .5" strip and pull it around. Amazing stuff. Probably going to get a dispenser or build one because snipping those pieces while working is a bit of a pain after 10 dishes.
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FractalXplora
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Registered: 02/11/06
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Loc: UK
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need some more parafilm, anyone wanna do me a deal on a nice strip to the UK, will hit up ya paypal of course. Cheers for the good advice all, gotta love messing with agar!
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hamloaf
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Registered: 12/23/09
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HL
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Base Icks
Registered: 03/19/10
Posts: 6,191
Loc: Shroomshire
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Quote:
FractalXplora said: need some more parafilm, anyone wanna do me a deal on a nice strip to the UK, will hit up ya paypal of course. Cheers for the good advice all, gotta love messing with agar!
Andymc just put a link up in the uk thread in the last couple of weeks for the para. Not to bad of price for a biiig roll of it. I lucked out and found a small piece on ebay for a fiver, should last me a little while till I can get a big roll of my own.
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Javadog
Continuing along
Registered: 05/03/10
Posts: 7,385
Loc: USA
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Quote:
FractalXplora said:
Quote:
No need to spend a wad of your parafilm wrapping them individually. Save your film for when you go to use them. Just use Glad-Wrap after pouring (while hot if you like) then put your sleeve over the stack then rubber band/clip/seal. Wrap the whole column solid then just cut the dishes out as you need them. That way they stay 'individually wrapped' until you need them.
Sounds like a plan
Indeed. An excellent point. Uninoculated Agar does not need GE. ;0)
-------------------- Boyd Rice told my brother that life is a corny pack of freesakes Myco-tek.org
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Mephistophelian
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Registered: 08/14/08
Posts: 2,527
Loc: Camp Crystal Lake
Last seen: 1 year, 27 days
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Re: Potato Dextrose Agar (PDA)! [Re: Javadog]
#13029355 - 08/09/10 10:08 PM (13 years, 7 months ago) |
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Well you are correct about that.
But Glad-Wrap does provide GE just fine as well. Been using it for 2 years up until I recently switched to Parafilm (there was a really good deal opportunity). Don't want people to get the wrong idea.
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