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Takeatrip
Stranger
Registered: 09/10/23
Posts: 183
Last seen: 2 hours, 6 minutes
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Contamination with drops on agar or loop method
#28648271 - 02/05/24 10:44 PM (3 months, 9 days ago) |
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How come when I try putting spores onto agar it often goes an off white colour and doesn’t germinate a thing with some syringes? I even get this with streaking with a loop? You can see where I went with the loop but no growth. I have had this with different vendors too. I do flame sterilise the loop beforehand and shake the syringe and the spores are visible. I have had some luck using BRF cakes and transferring from them onto agar but even that recently has not worked out. I am keeping everything as sterile as possible but just not getting much luck and wasting dishes.
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WyoMX

Registered: 07/06/15
Posts: 2,232
Loc: PNW
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Re: Contamination with drops on agar or loop method [Re: Takeatrip] 1
#28648276 - 02/05/24 11:00 PM (3 months, 9 days ago) |
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Whats your agar recipe? If your able to germinate on brf then not getting growth on agar something is up it seems like.
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Takeatrip
Stranger
Registered: 09/10/23
Posts: 183
Last seen: 2 hours, 6 minutes
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Re: Contamination with drops on agar or loop method [Re: WyoMX]
#28648953 - 02/06/24 12:32 PM (3 months, 8 days ago) |
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Quote:
WyoMX said: Whats your agar recipe? If your able to germinate on brf then not getting growth on agar something is up it seems like.
I buy them in from a company that makes them, they are very good and I am pleased with transfers I have done on them. They are Malt Extract Agar plates. I just find it odd that only one strain has been successful when using spores straight to agar, the rest have been unsuccessful. I have just prepared some more so I will see how those go from a new vendor. I am also doing some BRF cakes as a back up for germination.
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HappinessStan
Fungivore



Registered: 10/10/12
Posts: 1,762
Loc: Worcester, UK
Last seen: 4 days, 9 hours
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Re: Contamination with drops on agar or loop method [Re: Takeatrip] 1
#28648970 - 02/06/24 12:42 PM (3 months, 8 days ago) |
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You can simply make a brf/verm puck in a jar with a gas exchange filter and shoot spore syringes onto that, then pluck clean myc to agar. Really helps with spore syringes. I've always found it hard to get a decent germination with spore syringe to agar due to the excess water. Brf pucks work brilliantly and you can even let them grow out and clone from them.
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joze



Registered: 11/10/20
Posts: 1,056
Loc: PNW
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Re: Contamination with drops on agar or loop method [Re: HappinessStan]
#28649191 - 02/06/24 04:12 PM (3 months, 8 days ago) |
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When you don't see any fuzzy mycelium, and instead the agar turns a milky white color, that's usually a sign of bacterial contamination.
How are you introducing the spores to the agar? You mentioned a flame-sterilized loop. Are you letting your loop cool down for a few seconds before you wipe up some spores? If it's still red hot you'll just kill the spores you touch.
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