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NightPuma1
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Registered: 03/28/16
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Going Agar and just want to verify a few minor things...
#23866216 - 11/25/16 02:42 PM (7 years, 2 months ago) |
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Ok so I'm getting into doing Agar. I have a couple of questions that I have not been easily able to find the answers to so if someone is so kind...
1. In the video I've seen, the person took the ENTIRE piece of agar from the petri dish with healthy mycelium growing on it and transferred it into a sterile Wild Bird Seed (WBS) jar. I sort of though that you would slice it into several pieces and transfer those pieces to several jars. Is a whole petri dish worth of agar put into a jar overkill or am I mistaken?
2. Also, regardless if you transfer an entire petri dish worth of agar into a WBS jar or not, shouldn't you cut it up so that there are more points of contact in the WBS jar for better inoculation?
3. A lot of the agar I am finding people talking about is agar powder. However, I also know it comes in sort of "gelatinous sticks" which you need to grind down and that is sort of what I am leaning towards. The main reason I want to go for the sticks is because I want to use this extremely easy tek called the Grocery Store Agar Tek by Helltick that uses the fewest number of ingredients I've come across and also seems to be the cheapest:
https://www.shroomery.org/9427/Grocery-Store-Agar-Tek
Anyone have any problems with this?
Thank you for the help!
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Ferather
Mycological



Registered: 03/19/15
Posts: 6,325
Loc: United Kingdom
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Re: Going Agar and just want to verify a few minor things... [Re: NightPuma1]
#23866237 - 11/25/16 02:49 PM (7 years, 2 months ago) |
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Quote:
NightPuma1 said: 1. In the video I've seen, the person took the ENTIRE piece of agar from the petri dish with healthy mycelium growing on it and transferred it into a sterile Wild Bird Seed (WBS) jar. I sort of though that you would slice it into several pieces and transfer those pieces to several jars. Is a whole petri dish worth of agar put into a jar overkill or am I mistaken?
That is a weird and unusual thing to do, not overkill but incorrect.
Quote:
NightPuma1 said: 2. Also, regardless if you transfer an entire petri dish worth of agar into a WBS jar or not, shouldn't you cut it up so that there are more points of contact in the WBS jar for better inoculation?
Yes you should cut it up for that exact reason, and to isolate clean areas, or general isolation, well done.
Quote:
NightPuma1 said: 3. A lot of the agar I am finding people talking about is agar powder. However, I also know it comes in sort of "gelatinous sticks" which you need to grind down and that is sort of what I am leaning towards. The main reason I want to go for the sticks is because I want to use this extremely easy tek called the Grocery Store Agar Tek by Helltick that uses the fewest number of ingredients I've come across and also seems to be the cheapest:
https://www.shroomery.org/9427/Grocery-Store-Agar-Tek
Anyone have any problems with this?
Thank you for the help!
Can you provide a link or image for these "gelatinous sticks" thanks.
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enlightenment
alchemist


Registered: 08/09/09
Posts: 1,647
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Re: Going Agar and just want to verify a few minor things... [Re: Ferather]
#23866248 - 11/25/16 02:53 PM (7 years, 2 months ago) |
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I often use 1/4 to 1/2 of a petri dish to inoculate a jar. Why? Because it colonizes faster. I use the colonized jar for g2g.
The amount of agar does not matter. The smaller the longer to colonize of course.
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Pastywhyte
Say hello to my little friend



Registered: 09/15/12
Posts: 37,810
Loc: Canada
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Re: Going Agar and just want to verify a few minor things... [Re: Ferather]
#23866255 - 11/25/16 02:56 PM (7 years, 2 months ago) |
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1. The bigger the wedge the faster the colonization. You can use a small wedge if you want but it will take longer.
2. Cutting the wedge up does provide more knock points but it's also extra exposure time. My tic tac toe agar wedge drop helps cut down the exposure and results in 9 wedges per jar for very fast colonziation.
https://www.shroomery.org/forums/showflat.php/Number/22601023
3. I used the sticks back when I did the grocery store tek. It's fine to use but powder dissolves better and is less prep. Both work tho. The grocery store tek is a bit dated but still works. Few things I would do differently myself.
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Ferather
Mycological



Registered: 03/19/15
Posts: 6,325
Loc: United Kingdom
Last seen: 1 year, 2 months
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Re: Going Agar and just want to verify a few minor things... [Re: Pastywhyte]
#23866326 - 11/25/16 03:18 PM (7 years, 2 months ago) |
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Pretty similar to your high spawn ratio approach
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NightPuma1
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Re: Going Agar and just want to verify a few minor things... [Re: Ferather]
#23867462 - 11/25/16 10:11 PM (7 years, 2 months ago) |
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The link is right there in my OP.
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Boogieman47
Let's boogie


Registered: 03/05/16
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Re: Going Agar and just want to verify a few minor things... [Re: NightPuma1]
#23867513 - 11/25/16 10:31 PM (7 years, 2 months ago) |
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I use those sticks all the time they are cheap and works the same if you get a coffee grinder you can gdt them almost to a powder ... and i make my agar softer and it pretty much breaks up every where on its own if i do a tiger drop but if i want to make more jars ill either cut 4 wedges or do an li with the agar
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NightPuma1
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Re: Going Agar and just want to verify a few minor things... [Re: Pastywhyte]
#23867780 - 11/26/16 01:48 AM (7 years, 2 months ago) |
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Ok here's my final question:
Is it better to grow a bunch of petri dishes with healthy mycelium and then transfer them directly to say 16-20 jars of WBS or should you use just a few petri dishes to inoculate 3-4 jars of WBS and THEN do GTG transfers to inoculate the the 16-20 jars?
I sort of see there being a little more advantage in growing out say 30 petri dishes and using only the healthy ones to transfer to the 16-20 jars of WBS because with GTG transfer you never really know if contaminants got into your master jar from one transfer to another since there are several.
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enlightenment
alchemist


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Re: Going Agar and just want to verify a few minor things... [Re: NightPuma1]
#23867842 - 11/26/16 03:15 AM (7 years, 2 months ago) |
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If you just want to get a lot of grain in short time you should use a few plates to check for healthy mycelium and then G2G. This is faster and easier.
The advantage you mentioned is true. But if you have a good sterile technique G2G is a good way to expand grain very fast.
+ You have the same "genetic pool" in the g2g jars.
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NightPuma1
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Re: Going Agar and just want to verify a few minor things... [Re: enlightenment]
#23868642 - 11/26/16 11:16 AM (7 years, 2 months ago) |
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So if my ultimate goal was to get 16-20 WBS jars fully colonized, how many petri dishes would you recommend I start with? I.E. if I started with a spore syringe and 12 petri dishes, I feel like there is a pretty solid chance I'd definitely be able to clean up at minimum 3-4 of them with healthy mycelium. Then what? I should use just only 1 of those 3-4 healthy ones to inoculate a jar of WBS. Then wait until that jar is fully colonized, and then GTG transfer that ONE master jar into 16-20 sterilized jars?
I guess my one issue there is that I can't fit 16-20 jars in my glovebox at once to GTG transfer them all at once - the most I would usually be able to fit in my glovebox at once would be like 6-7 plus the master jar.
That means I would need to do 3 runs to get all 16-20 which means opening the glovebox 3 times. This is why in the past I would usually make 3 master jars and use one per run, as opposed to using the same master jar for all 3 runs because as soon as I open the lid on my glovebox unclean air rushes in. Even if I kept the lid on on the master jar, I feel like this is a pretty big risk of contaminants getting in.
How do you recommend I proceed? Appreciate you.
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Munchauzen


Registered: 06/22/11
Posts: 14,342
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Re: Going Agar and just want to verify a few minor things... [Re: NightPuma1]
#23868653 - 11/26/16 11:20 AM (7 years, 2 months ago) |
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Quote:
NightPuma1 said: So if my ultimate goal was to get 16-20 WBS jars fully colonized, how many petri dishes would you recommend I start with? I.E. if I started with a spore syringe and 12 petri dishes, I feel like there is a pretty solid chance I'd definitely be able to clean up at minimum 3-4 of them with healthy mycelium. Then what? I should use just only 1 of those 3-4 healthy ones to inoculate a jar of WBS. Then wait until that jar is fully colonized, and then GTG transfer that ONE master jar into 16-20 sterilized jars?
I guess my one issue there is that I can't fit 16-20 jars in my glovebox at once to GTG transfer them all at once - the most I would usually be able to fit in my glovebox at once would be like 6-7 plus the master jar.
That means I would need to do 3 runs to get all 16-20 which means opening the glovebox 3 times. This is why in the past I would usually make 3 master jars and use one per run, as opposed to using the same master jar for all 3 runs because as soon as I open the lid on my glovebox unclean air rushes in. Even if I kept the lid on on the master jar, I feel like this is a pretty big risk of contaminants getting in.
How do you recommend I proceed? Appreciate you.
blenderless li in my signature
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NightPuma1
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Re: Going Agar and just want to verify a few minor things... [Re: Munchauzen]
#23868824 - 11/26/16 12:14 PM (7 years, 2 months ago) |
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Thank you for your response. I do feel as if the problem I asked about would still remain if I used a blenderless LI:
whether it was a petri dish with healthy mycelium, a fully colonized master WBS jar for GTG transfer OR a blenderless LI, it seems as though I still have to do something like 3 runs in order to inoculate 16-20 WBS jars just due to only being able to fit so many jars in at once. Whichever of the 3 options I choose I will still be limited by the number of jars I can fit in my glovebox at once, and this results in opening the Glovebox which increases the risk of contamination.
I guess the advantage I see in using a blenderless LI is that mixing the healthy mycelium agar with water creates "more" of that specific strain than what I would have had with just the agar plate.
However, this would mean I would NOT be doing any GTG transfers at all. Not that I have a problem with this, but people seem to swear by them. I guess with using a blenderless LI it would skip the weeks needed for the master jar to colonize.
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Munchauzen


Registered: 06/22/11
Posts: 14,342
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Re: Going Agar and just want to verify a few minor things... [Re: NightPuma1]
#23868858 - 11/26/16 12:29 PM (7 years, 2 months ago) |
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you should be using a still air box, not a glovebox. as for your issue the only thing i can tell ya is buy a bigger tote. i have a 106 qt tote and can fit 10 grain jars, 4 water jars, and 4 plates for the entire session.
its fine to briefly open the SAB to remove the jars to shake them. the SAB works by having wet walls, thats the most important factor, not letting it settle out.
yes LI means not having to do G2G. neither is superior, they are just different methods. doing g2g isnt some cultivation magic trick, its just a method for creating spawn like LI.
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NightPuma1
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Re: Going Agar and just want to verify a few minor things... [Re: Munchauzen]
#23871027 - 11/27/16 02:18 AM (7 years, 2 months ago) |
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Hey man thanks for the help - you'll be happy to hear I've been watching your videos and intend on actually listening to your advice.
One question (I know I'm sorry): in your video you use 2 grams of agar powder. If I instead bought it in chunk form (it came in two sticks pre-dyed green with food color), do I just grind up 2 grams worth or should I adjust the recipe in some way?
Also, does it have to be ground to complete powder or will the boiling help break it down?
Thanks.
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filthyknees
no coincidence


Registered: 03/08/13
Posts: 6,283
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Re: Going Agar and just want to verify a few minor things... [Re: NightPuma1]
#23871164 - 11/27/16 06:04 AM (7 years, 2 months ago) |
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Here's what I'd do.. I'd start with germinating spores on five plates, take at least two transfers to new plates before transferring to grains.
You can use three transfers per plate before the final plate used to innoc to save space. So if two/five plates germinate take transfers from the leading edge to new plates. If I wanted 20 grain masters I want to end up with 20 clean petris so I will make maybe six transfers from germinating plates leading edges (on to two plates)
Then after a few days transfer the leading edge again this time each transfer gets its own dish in prep to go to grain so long as growth looks clean. so five transfers from each of the previous six transfers = 30 plates, assuming ten go bad because you've never used agar before you have your 20 plates.
I'd aim for more than what youd think and not put all eggs in one basket. This is why I like one plate -> one jar. If you mess up and only have 10 good plates at the end, you can definitly do one plate -> two jars no problems.
If you want non-grain master jars, I would follow enlightenment & get a few grain masters and g2g them x10
For agar recipie you want it as ground as possible. All premix agar comes in powder form, so no boiling is required. I would try that.
-------------------- But if you're in a hurry, and really got to go If you're in a hurry, might have to find out slow That it's one thing to try and another to fly You get there quicker just a step at a time It's one thing to bark, another to bite The show ain't over till you pack up at night
Edited by filthyknees (11/27/16 06:10 AM)
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NightPuma1
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Re: Going Agar and just want to verify a few minor things... [Re: filthyknees]
#23871771 - 11/27/16 11:35 AM (7 years, 2 months ago) |
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Okay good but will the amount of agar change? I.e. 2 grams of powdered agar vs. 2 grams of agar sticks ground up or should I change the amount at all?
Thanks.
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filthyknees
no coincidence


Registered: 03/08/13
Posts: 6,283
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Re: Going Agar and just want to verify a few minor things... [Re: NightPuma1]
#23871773 - 11/27/16 11:36 AM (7 years, 2 months ago) |
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Powder or not 2g is 2g. Measurement doesn't change
-------------------- But if you're in a hurry, and really got to go If you're in a hurry, might have to find out slow That it's one thing to try and another to fly You get there quicker just a step at a time It's one thing to bark, another to bite The show ain't over till you pack up at night
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