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North Spore Shop: Injection Grain Bag

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Invisibleyutaka

Registered: 06/12/10
Posts: 544
First time pouring agar - what was your contamination rate?
    #14289760 - 04/14/11 12:15 PM (12 years, 9 months ago)

I poured a couple of PDA dishes the other day.  I poured a case of half pint wide mouth jars with the metal lids and bands as my petri dishes have not come in yet.  I had a 50% contam rate while working in my glove box and my left over agar is unscathed by contaminants. I'm working on better sterile technique and also closing up my arm holes some as they are too big.

First I sterilized the half pint jars for an hour at 15lbs.  I let them cool and seal.  Then I made my ager from the standard PDA recipe: 1L water, 200g potato broth, 20g agar, 20g dextrose scaled down to a 750ml bottle.  Working in the glove box is a little difficult.  I wiped the glove box out, placed my jars in the box, the glove box in a small room without ventilation and waited 10 minutes.  At this point the dishes were stacked four stacks of three.  I had wiped them down prior to placing them in the box, I probably should have wiped them down with IPA just before pouring while inside.  I put on gloves and tyvek sleeves, sanitized with alcohol and then got to pouring.  I left the top off of my agar bottle for the duration of the pour.  I then went through the stacks taking the band off, then the lid, pouring, then sealing back up with the lid and the band.

I probably would have done better to remove the bands on all the jars prior to pouring and then sanitize, pour, place the lids on and then after pouring all the dishes, sealing them back up with the band.

I feel this was a successful pour as it was my first and I didn't trash all of them.  The only contaminant on the plates was thin, wispy cobweb mold.

Enough about me...  what was your contamination rate on your first agar pour?
What was your contamination rate on your first agar pour?
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What type of agar did you pour with?
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About how many plates did you pour?
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North Spore Shop: Injection Grain Bag


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