Online agar sales

[El Presidente Hillbillios]

Hey guys,

I saw this and thought some of you guys might be interested. Not everyone can get to an Asian supermarket.

http://www.asiangrocerystore.com.au/swallow-globe-agar-agar-powder.html

[NSF]

Hey guys,

I saw this and thought some of you guys might be interested. Not everyone can get to an Asian supermarket.

http://www.asiangrocerystore.com.au/swallow-globe-agar-agar-powder.html

 

Thanks for the link Hillbillios.

Has anyone used this brand? I've got a packet of it sitting on my shelf and my first petri dishes were a complete disaster (no growth at all from a tissue sample, due primarily to me keeping them in with the hot water service, not having a PC and not mixing in the right nutrients. I guess it's really due to not having the right knowledge to begin with.)

So I'd just like to know it was the user and not the agar. But I'm pretty certain I know the answer already.

On that topic, when Growing Gourmet and Medicinal Mushrooms specifies the different growth parameters, which is the petri dish incubation temperature? Under the heading of Spawn Run? That doesn't read right.

Edited January 25, 2011 by NSF

[Magicdirt]

Thanks for the link Hillbillios.

Has anyone used this brand? I've got a packet of it sitting on my shelf and my first petri dishes were a complete disaster (no growth at all from a tissue sample, due primarily to me keeping them in with the hot water service, not having a PC and not mixing in the right nutrients. I guess it's really due to not having the right knowledge to begin with.)

So I'd just like to know it was the user and not the agar. But I'm pretty certain I know the answer already.

On that topic, when Growing Gourmet and Medicinal Mushrooms specifies the different growth parameters, which is the petri dish incubation temperature? Under the heading of Spawn Run? That doesn't read right.

 

I've never used that brand, but you should get some growth on agar even if your sterile technique is not up scratch. You'd be more likely to grow contaminants than mycelium though, either that or contaminated mycelium.

I've used straight agar before and it worked OK but you really need some nutrient source to get good growth.

In GGMM when they speak of the spawn run they are already a few steps past the stage where petri dish cultures are being discussed.

The spawn run is usually when colonised grain spawn is mixed with a bulk substrate and the spawn "runs" through the substrate.The incubation temps are similar for the spawn run and for agar culture but a few species require slightly different temps.

Chapter 12 covers petri dish culture, but for species specific incubation temps just google it.

[NSF]

Ok thanks for that. One more hijack...i put some cooled boiled water into some boiled jars (pasta sauce) then added some agar (and brown sugar and flour just cause) then i put the lids on loosely and boiled the jars standing in a covered pot for an hour. Is this the go?

After 3 weeks in the dishes in near the hot water service some agar had split, some had curled. I was away so don't know whether anything grew in the three weeks but there were few traces of anything at the end.

How long should agar in a dish stay viable for?

[Magicdirt]

It sounds like the agar dried out and the culture died, possibly it was too hot.

There's really no need to incubate agar cultures in summer, a closet in a cooler part of the house would be better, if it gets too hot the cultures will contaminate.

In winter you may need incubation but I found the cooler temps keeps them growing slowly so you don't get a snowball effect and end up with too many cultures to maintain.

Make your agar up in a saucepan first and pour it into jars before it cools, it can be kept in the fridge for a few months like that until you're ready to sterilise in the microwave if you don't have a Pressure Cooker.

Agar can be microwaved in a taco sauce jar with the lid on(strangely it doesn't spark or cause problems). Leave the lids slightly loose and tighten them as soon as the microwave stops, but use a glove or a tea towel as they will be very hot. just boil it and keep it hot for a few minutes, no need for thermometers just keep it hot for at least 2,preferably 3 minutes, you may have to start and stop the microwave so it doesn't overheat or turn the power down once it has boiled to keep it close to boiling point. If it boils too hard it will dehydrate so keep it at the point where it only just boils

Here's an old post with some excellent agar recipes, take your pick

Agar recipes

[NSF]

A huge many thanks Magicdirt!

So many questions answered before I even needed to ask them. Thank you for cutting me off at the pass. That right there is some really useful guidance!

I'm curious though, if you're making agar in a non sterile environment, like you suggest, then storing it in the fridge why does nothing grow? Oh and do you cook the agar? As in, teaspon of vegmite, teaspoon of agar, 1 cup of water, heat it just until the agar dissolves or do I boil the beejesus out of it as a pasteurising/sterilising tek?

Also, what temp do you think is good to pour it at after microwaving? (not too specific but should I be able to handle the jar with bare hands? Pour when it's steaming hot?) I guess, if it's hot then it could kill anything in the petri dish as it gets poured in.

Cheers,

Andrew

[MagikF]

If you want to use Asian agar, get the one without coloring or additives such as sugar or flavoring. There are brands out there that are just straight agar.

The Swallow one I think is from Indonesia. They make a packet that is just straight agar. You can also get one's from Thailand such as Telephone brand.

[Magicdirt]

A huge many thanks Magicdirt!

So many questions answered before I even needed to ask them. Thank you for cutting me off at the pass. That right there is some really useful guidance!

I'm curious though, if you're making agar in a non sterile environment, like you suggest, then storing it in the fridge why does nothing grow? Oh and do you cook the agar? As in, teaspon of vegmite, teaspoon of agar, 1 cup of water, heat it just until the agar dissolves or do I boil the beejesus out of it as a pasteurising/sterilising tek?

Also, what temp do you think is good to pour it at after microwaving? (not too specific but should I be able to handle the jar with bare hands? Pour when it's steaming hot?) I guess, if it's hot then it could kill anything in the petri dish as it gets poured in.

Cheers,

Andrew

 

If you pour agar in a non sterile environment you will have contaminants grow on the agar, the cool temp in the fridge will slow the growth but it will still have contaminants. That's why it needs to be sterilised prior to use.

When you make the agar mix you will have to boil the water for the agar to completely dissolve, I boil it dissolve the agar and when that's done I add the other ingredients while it's simmering. This step is not considered sterilisation it's just to mix the ingredients into a suspension.

You don't need a thermometer unless you're adding peroxide, just pour while it is still liquid. Use a glove though as it will be too hot to handle. If you are pouring a larger batch -like a litre you have a bit more time before the agar sets because of the thermal mass of the liquid so you can let the agar cool until it's just cool enough to handle and then pour it, agar poured at a cooler temp will be less likely to cause condensation in the petri dishes too, but for a smaller batch just pour it quick.

Many will disagree but I think agar should only be poured in front of a flow hood - there's just to much chance to get contamination at that stage.

If you intend to add peroxide you'll need a thermometer as peroxide breaks down at around 60 deg Celsius , so you have to wait for the agar to cool to that temp to add the peroxide.

It can be tricky to get the peroxide mixed in and the agar poured before the agar solidifies though. the peroxide is added after sterilisation.

If you don't have a sterile environment taco sauce jars are convenient because the agar can be sterilised in the jar and you don't need to pour it and expose it to contamination.The taco sauce jar becomes the petri dish and the lid is only opened prior to inoculation, bypassing the pouring stage and eliminating the possibility of contamination at that stage.That's how I used to do my agar work and my contam rate was less than 3%. I used to PC the jars at first but the microwave method worked just as well.

You should have a glove box for your inoculation work, open air inoculation work for some but I never had much success. Pouring agar in a glovebox is a real bitch though, the box steams up and you cant see things and trying to handle hot stuff in a box where your movement is restricted is very tedious, so that's another reason I used to use taco sauce jars.

[El Presidente Hillbillios]

I have been using agar for a while and dont have a flow hood or even glove box.

All i do is mix up my malt extract agar (10 grams of light malt extract and 9 grams of agar) With 500ml of just boiled water. Then i microwave it for a couple of mins on low so is just simmers.

I take the mix out with a glove and pour it immediately into The short mexican dip jars that have been fitted with a self healing injection port. This is a jar that has been used to make sterile water, but just to show the jars i use.

The jars are then sterilised in a preasure cooker fro 20-30 mins with the lids on loose. As soon as they finish i take them out and close the lids.

Now you have sterile agar in its own little sterile environment. Thens its as easy as injecting spores or whatever you want through the port at the top, be carefull though because the jars are under vacuum at this point.

My next step is then to wait for the myc to grow and then with a long 16g needle take a little plug of colonised agar. Again through the injection port without opening the jar. then this can either start another agar jar or a liquid culture.

[Magicdirt]

Cool tek man!

If you inject spores into something under a vacuum it can suck the contents of the syringe into the jar.

To equalise the vacuum make an airport syringe and poke it through the injection port.

Some people glue syringe filters on the lids so the culture can breathe and it prevents the vacuum, but I prefer to just use the airport syringe.

To make an airport syringe just pull the plunger out of a syringe and fill it with polyfill, then cover the open end with micropore tape.

You only have to sterilise it once, but the needle needs to be sterilised before each use.

[NSF]

@hillbilly: great tek...where's a good place to source injection ports?

Edit: i just saw your other thread about these, so i'll wait and see what turns up.

Also, just how much agar can you suck up with the syringe? Doesn't the gel clog the needle?

Edited January 30, 2011 by NSF

[El Presidente Hillbillios]

Yeah to eqalise the vacumm i just let it breath through a needle with a syringe filter.

I just take a plug, by this i mean have some air in the syringe, just stab the agar and draw in a alittle. Then once in the lc jar just blow it out with the air in the syringe. so i dont actually suck up any agar, just kinda take a core with the needle.

[El Presidente Hillbillios]

yeah i tried adding filter to let these breath, but i find way less contams if i just leave em sealed.

My thinking is there is plenty of oxygen in there to get a full plate grown.

I even use the same sealed jar idea for liquid culture

[NSF]

I just take a plug, by this i mean have some air in the syringe, just stab the agar and draw in a alittle. Then once in the lc jar just blow it out with the air in the syringe. so i dont actually suck up any agar, just kinda take a core with the needle.

 

So it's a bit like a mini spud gun? You just take a little core? But do you take several cores without removing the needle from the jar? Or do you take a plug, spit it into a grain spawn jar, flame sterilise and back into the salsa jar?

How many 'cores' do you use to colonise some grain or rice flour?

[El Presidente Hillbillios]

I would normaly do just one core into a lc. then i use this as my master lc to start more lc's. Then i use the Lc's to start grains.

That agar then stays in the fridge with the original master lc

[migraineur]

If you want ready made good quality agar then go to http://www.mycosupply.com and buy yourself some potato dextrose agar or malt extract agar. It's good quality and it saves you the hassle of boiling potatoes and making your own mix etc.

They also have some other cool stuff for your agar work like metal inoculation loops and a mushroom cloning kit.

[NSF]

(10 grams of light malt extract and 9 grams of agar)

 

I know that if I listened to migraineur then I wouldn't need to ask this question but what the hey!

light malt extract...this is the home brew ingredient isn't it? http://www.coopers.com.au/corporate/malt-extract

Yes, that could be the stupidest thing I've asked out loud.