Zakkery said:

Picture tax. Are the little darker spots in the mycelium contaminants? They appear as the Myc grows out so I don't think it's little LME nuggets.

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covertjoy said:

40 plates poured, 4 different recipes (10 of each).

Recipe 1 (bottom left)

200ml distilled water
3.0g agar
3.0g light malt extract

Recipe 2 (top left)

200ml distilled water
3.0g agar
3.0g casein hydrolysate

Recipe 3 (bottom right)

200ml distilled water
3.0g agar
3.0g light malt extract
0.1g nutritional yeast

Recipe 4 (top right)

200ml distilled water
3.0g agar
3.0g casein hydrolysate
0.1g nutritional yeast

It's an experiment. I have 2 clones, 6 various transfers and 2 germinations to do. For each inoculation I will repeat 4 times across each of the different recipes. This way I will be able to compare like for like for each of the recipes in different situations.

I poured them a little hotter than I usually do due to time constraints so there is probably a fair amount of condensation. I'll leave them in front of the flow hood until tomorrow and do the inoculations at about +20 hours.

Initial observations are that light malt extract is more orange where as the casein hydrolysate is very pale. Nutritional yeast makes the recipe cloudy. Casein hydrolysate is much nicer to work with than malt extract.

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covertjoy said:

Just made spore suspensions for:

• Ps. Hispanica
  
• Ps. Cubensis (Stargazer) × Ps. Natalensis [F1]
  

Ps. Hispanica is from a spore print.



This is the second of two Hispanica spore prints I had. The first one I failed to get any germinations from so am having another shot at it. I dropped the spore print into 2ml of sterile distilled water in a test tube with a steel ball bearing. I shook and vortexed the tube then used a sterile pipette to transfer the suspension into another test tube.

The spore print is on card and when shaking it (less so vortexing) it decimated the card. When I was counting the spore concentration with a haemocytometer and microscope it was very difficult due to there being so much paper debris. Perhaps this will affect germination in some way. The card may contain toxic glues. Please don't use card for spore prints. The spore concentration is 1,875.0 single spores per drop (assuming 0.05ml per drop) with an additional 0.33 clusters of spores per single spore.

Inoculated 4 petridishes of different recipes each with 5 drops of the spore suspension in a crosshair formation. If I get any successful germination then I will do a serial dilution and inoculate more plates to try to isolate a monokaryotic culture.



Ps. Cubensis (Stargazer) × Ps. Natalensis [F1] used a very sparse looking swab. I pulled the end of the swab away from the wooden stick using tweezers and snipped it into 3ml of sterile distilled water in a test tube with a steel ball bearing. I shook and vortexed the tube and then used a sterile pipette to transfer the suspension into another test tube. In the final test tube there is about 2.25ml of suspension since the swab absorbed some of the liquid.

The spore solution for this one was much cleaner, I like working with swabs, and the concentration was higher than I expected. There are 5,062.5 single spores per drop (assuming 0.05ml per drop) with an additional 0.23 clusters of spores per single spore.

Inoculated 4 petridishes of different recipes each with 1 drops of the spore suspension mostly just to test it's germination and potential contamination rate. I'm planning to send the spore suspension directly to spawn for top fruiting to select an F2.

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jungatheart said:

BeefSupremeJr said:

been getting so much shit for this in my disco i had to come post it.  laugh it up, fuzzballs, but this is the ideal media bottle setup. you may not like it but this is what peak performance looks like.

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I like it! Where can I get one?

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applesmasher420 said:

Zakkery said:

Picture tax. Are the little darker spots in the mycelium contaminants? They appear as the Myc grows out so I don't think it's little LME nuggets.

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Its hard for me to tell but it may be the beginning of hyphal nots / primodia. The mycelium gets real dense where it is going to try to fruit.

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I've been seeing something similar in some of my Pan cyan plates -- looks like great, clean growth, then a little dense mycelial bump shows up. I'd been assuming mold, but I never opened the plate after transfer?

BeefSupremeJr said:

jungatheart said:

BeefSupremeJr said:

been getting so much shit for this in my disco i had to come post it.  laugh it up, fuzzballs, but this is the ideal media bottle setup. you may not like it but this is what peak performance looks like.

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I like it! Where can I get one?

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the zon.

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covertjoy said:

https://www.amazon.co.uk/NRS-Healthcare-Handle-Eligible-relief/dp/B00FRGQ77W

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CaptainPuffy said:

I'm trying to clean up a bunch of strains using agar. Mostly dealing with bacteria. Trying to use water agar. Using a SAB and typical sanitization routines. If anyone has any advice, I'd appreciate it.

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Pnin said:

Does it? I threw out like four JF plates I made testing a syringe because they all looked bunk and nothing like other myc I had personally seen.

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BeefSupremeJr said:

yea i had that experience. fuzz bucket.  someone told me it always looks like that.  lame strain if you ask me..

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Quote:

Where are the strains at the moment? I.e. Where will your first transfer be from?

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