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small scale agar work

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#1 teesus



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Posted 01 December 2008 - 11:32 AM

i want to show you my agar process
it is very low-fi but it works and i post it for those who never gave agar a try and think it is complicated. if you have a bit of practice it is not complicated at all. its a cheap and easy way to germinate spores, do cloning or sub strain selecting.
there are lots of other methods for each of this steps which work great, you might find improvements that work better for you. if you have a glove box you can use that for inoculation. i do my agar work open air in a kitchen, thats why i have to take care a bit more.

i am using glas petri dishes, agar from orient food store and malt extract.
first step is preparing of the agar. this is very easy. for 200 ml of water 4 g agar powder and 4 g of malt extract. boil the water and put agar and malt in. stir until all is dissolved
next is pouring the plates. i fill the agar in to a height of 3 or 4 mm.
before sterilization i put the plates on stables and wrap them with aluminium foil. it helps to keep them clean after they leave the PC, but i do it mainly to stabilize them in the PC during sterilization. i wrap the tools for inoculation in foil too and PC them with the plates.
PC is running for 20 min. while the PC is hot the agar is liquid, so its very important that the PC is not disturbed till it has cooled down and the agar is solid. if hot agar spills over the plate stables the batch might be ruined.

as the plates are steril and cooled down its time for inoculation. i am doing this work next to a burning gas stove. first i wash my hands and place the plate stable 20 cm next to the flame. you can't see the flame in the pictures which sucks, but it is burning. to explain the flame thing i painted this little pic.
the flame creates a stream of hot air that distracts airborne stuff from landing on your plate while you're working. this tek works well with a bunsen burner. Due to the fact that i'm using a stove, the area which is free from contamination is much smaller than it would be with a bunsen burner, i have to work near by the flame and be carefull not to burn my fingers or the spores.
i scrape a tiny bit of spores from a print with the inoculation needle and place it on the first plate of the stable, then the next and so on. i open the lid just a small gap and dip the needle with the spores on the agar. the faster this is done the smaller is the risk to catch a contamination.
when inoculation is done i seal the plates with wrap foil and store them upside down to prevent lid condensation.
thats all i do to germinate spores on agar. for tranfering agar to agar or agar to grain i am using the same method. i hope this is useful for some.
have a nice day!

Attached Thumbnails

  • inoc3.jpg
  • tools3.jpg
  • tower3.jpg

Edited by coorsmikey, 26 May 2016 - 05:00 AM.
Added tag

  • Lakegal7 likes this

#2 teesus



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Posted 01 December 2008 - 11:35 AM

two more pics of sealing the plates

Attached Thumbnails

  • sealing3.jpg

#3 karmakraze


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Posted 01 December 2008 - 05:25 PM

cut a chunk of saran wrap off to seel the petries good Idea I will have to borrow it thank you. Also ? What kind of contam rate if any do you have.


#4 teesus



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Posted 01 December 2008 - 05:47 PM

contams do happen. when i am working sloppy there is a contam rate of mabe 20%, but when its done carefully mostly all plates stay clean. one out of 10 is normal, so contam rate is 10%. to insure i always do some more plates than needed.
the wrap foil idea was from a grow book i had. it works very good. use a sharp knive and cut even trough the roll. if the cut is not even rolling up the foil is difficult.
  • Ferretious likes this

#5 prism


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Posted 01 December 2008 - 06:01 PM

Awesome tek!! now where did I put those plates?

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