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MysticPop
AlienInWonderland



Registered: 05/05/15
Posts: 132
Last seen: 2 months, 30 days
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Need advise on Inoculation with Agar Wedge Vs LI Vs LC
#21841570 - 06/22/15 12:48 PM (8 years, 7 months ago) |
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Hi All
I have come to know that there are 3 good different ways to inoculate grain jars. These are agar wedge, LI and LC. I have the following questions:-
1. Which is the safest method to do the inoculation? 2. Why is one preferred over the other? 3. Which has most success rate? 4. Any Recommended reading links etc?
I have some plates. I partly transferred the rhizophoric strains to new agar plates and i Have some left overs as well. I plan to inoculate the grains with these plates. Please refer to link http://www.shroomery.org/forums/showflat.php/Number/21840040 to see the plates pictures. I am also looking for advise and expert thoughts to know if these plates are good to do grain inoculation or not?
Cheers Mystic
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JvF
Fletcher Detcher


Registered: 02/13/14
Posts: 2,662
Loc: Chicago
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Re: Need advise on Inoculation with Agar Wedge Vs LI Vs LC [Re: MysticPop]
#21841619 - 06/22/15 01:02 PM (8 years, 7 months ago) |
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Agar wedge is definitely solid. Personally, i find it to be the most reliable, however it's slower than li and lc.
Wedge to lc is good, but contams are hard to spot. Plus it still has to be colonized, leaving more room for contams to grow. Many growers love LC, im not one of them.
LI is by far my favorite. Throw an agar wedge in a jar of sterile water and glass shards, shake, and inoculate. About the same speed as LC, but you know that the wedge was vlean and the water was sterile. Ive had more success with LI than LC, 100% success woth LI and about 70% with LC.
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MysticPop
AlienInWonderland



Registered: 05/05/15
Posts: 132
Last seen: 2 months, 30 days
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Re: Need advise on Inoculation with Agar Wedge Vs LI Vs LC [Re: JvF]
#21841631 - 06/22/15 01:06 PM (8 years, 7 months ago) |
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What LI Tek do you recommend? Also i heard we need a special blender for the LI. Do you know which blender isthe best? I dont want to make my own blender, but can buy one
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JvF
Fletcher Detcher


Registered: 02/13/14
Posts: 2,662
Loc: Chicago
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Re: Need advise on Inoculation with Agar Wedge Vs LI Vs LC [Re: MysticPop]
#21841815 - 06/22/15 02:16 PM (8 years, 7 months ago) |
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Ive seen the blender method and have always found it unnecessary.
You can either spray a bit of sterile water from a sterile syringe onto the plate, then suck it back up after agitating the myc with the needle
OR
What i like to do is sterilize a jar of water thats filled about 15%(if using a quart jar) with some shards of glass(this helps break up the myc). Then i take the colonized agar and place it into the water.
After that, agitate the mycelium by shaking vigorously, but not enough to hit the filter on top. The glass should knock a good anount into the water
Suck it into a syringe and inoculate or store. You could also store the jar as is
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MysticPop
AlienInWonderland



Registered: 05/05/15
Posts: 132
Last seen: 2 months, 30 days
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Re: Need advise on Inoculation with Agar Wedge Vs LI Vs LC [Re: JvF]
#21841849 - 06/22/15 02:29 PM (8 years, 7 months ago) |
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Do you have any tek on this method? I dont have Shards of glass available
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MysticPop
AlienInWonderland



Registered: 05/05/15
Posts: 132
Last seen: 2 months, 30 days
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Re: Need advise on Inoculation with Agar Wedge Vs LI Vs LC [Re: MysticPop]
#21841853 - 06/22/15 02:30 PM (8 years, 7 months ago) |
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Also do you drop the whole colonized agar or just a wedge? Which filter is there on top? Some pics or some tek would help a lot ..
Also thank you so much for your help
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Mad Season
hookers and blackjack



Registered: 09/16/12
Posts: 12,666
Loc: Canada
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Re: Need advise on Inoculation with Agar Wedge Vs LI Vs LC [Re: MysticPop]
#21841904 - 06/22/15 02:40 PM (8 years, 7 months ago) |
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All these things have to stem back to agar work. Personally I feel LC is stupid to do without agar. You inoculate multiple lcs with clean wedges, get them in syringes and test them on multiple plates. I'd even test LI on plates before using it. Testing liquids on agar plates before using them on grains is the best thing you can do, and has a 99% chance of making sure the liquid is clean. It may take a week or 2 of extra time, but afterwards you'll have lots of clean inoculant.
Agar wedge work is the best way to work on your sterile techniques though. The more plates you work with, the better you get. This is why I've been mainly doing a2gs. Tiger drop is the whole plate, you can even drop a small wedge. You just want to keep it clean and fast.
Edited by Mad Season (06/22/15 02:43 PM)
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maddchef
Vaginal escape artist



Registered: 09/04/09
Posts: 5,602
Loc: Your mom's vag
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Re: Need advise on Inoculation with Agar Wedge Vs LI Vs LC [Re: MysticPop]
#21841910 - 06/22/15 02:41 PM (8 years, 7 months ago) |
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You can skip the jar if you're just going to agitate.
Put a few ccs on a plate and "scratch" the plate with your sterile needle and suck the liquid back up. Now you have just a tiny LI.
Honestly if you're not making enough LI to get full coverage then its not worth the extra step, just drop a wedge in.
-------------------- In the land of the blind, the one eyed man is king. All mushrooms are edible, but some only once..... Easier than cakes I do science and shit.
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MysticPop
AlienInWonderland



Registered: 05/05/15
Posts: 132
Last seen: 2 months, 30 days
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Re: Need advise on Inoculation with Agar Wedge Vs LI Vs LC [Re: maddchef]
#21841943 - 06/22/15 02:49 PM (8 years, 7 months ago) |
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I want to fruit in 2 monotubs.. Currently I have 4 left over plates(See this http://www.shroomery.org/forums/showflat.php/Number/21840040). These are the plates i wish to inoculate the grain from. I made transfers to around 14 plates from 8 plates out of which the above 4 seemed to me a valid candidate for grain inoculation. Eventually i wish to have 2 monotubs full worth of material.. Also i want to get stuff growing.. not have had any growth yet ..
So questions :-
1. Are the plates shown here good for growth ? http://www.shroomery.org/forums/showflat.php/Number/21840040 2. How much growth can i expect from these.
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thoraxx
Wizard


Registered: 12/27/13
Posts: 580
Loc: Bavaria
Last seen: 4 years, 10 months
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Re: Need advise on Inoculation with Agar Wedge Vs LI Vs LC [Re: MysticPop]
#21841978 - 06/22/15 03:01 PM (8 years, 7 months ago) |
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Quote:
MysticPop said: I dont have Shards of glass available 
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Juiceh
Dabbing All Day



Registered: 09/25/12
Posts: 3,208
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Re: Need advise on Inoculation with Agar Wedge Vs LI Vs LC [Re: MysticPop]
#21842104 - 06/22/15 03:40 PM (8 years, 7 months ago) |
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Quote:
MysticPop said: Do you have any tek on this method? I dont have Shards of glass available 
Do your neighbors have windows?
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MysticPop
AlienInWonderland



Registered: 05/05/15
Posts: 132
Last seen: 2 months, 30 days
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Re: Need advise on Inoculation with Agar Wedge Vs LI Vs LC [Re: Juiceh]
#21842132 - 06/22/15 03:50 PM (8 years, 7 months ago) |
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LOL>> Not doing that..
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bodhisatta 
Smurf real estate agent


Registered: 04/30/13
Posts: 61,889
Loc: Milky way
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Re: Need advise on Inoculation with Agar Wedge Vs LI Vs LC [Re: MysticPop]
#21842180 - 06/22/15 04:06 PM (8 years, 7 months ago) |
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Break a pint glass or something or buy a soda in a glass bottle. If you can't figure out glass shards mycology is not going to be easy
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MysticPop
AlienInWonderland



Registered: 05/05/15
Posts: 132
Last seen: 2 months, 30 days
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Re: Need advise on Inoculation with Agar Wedge Vs LI Vs LC [Re: bodhisatta]
#21842205 - 06/22/15 04:14 PM (8 years, 7 months ago) |
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Glass shards can be arranged but the bigger question was,..which way to go :-
I want to fruit in 2 monotubs.. Currently I have 4 left over plates(See this http://www.shroomery.org/forums/showflat.php/Number/21840040). These are the plates i wish to inoculate the grain from. I made transfers to around 14 plates from 8 plates out of which the above 4 seemed to me a valid candidate for grain inoculation. Eventually i wish to have 2 monotubs full worth of material.. Also i want to get stuff growing.. not have had any growth yet ..
So questions :-
1. Are the plates shown here good for growth ? http://www.shroomery.org/forums/showflat.php/Number/21840040 2. How much growth can i expect from these.
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Grey
⇜ ✯ ⇝



Registered: 11/06/14
Posts: 6,223
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Re: Need advise on Inoculation with Agar Wedge Vs LI Vs LC [Re: bodhisatta]
#21842210 - 06/22/15 04:16 PM (8 years, 7 months ago) |
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Glass is pretty common.
I've squirted sterile water on agar plates with good success. Munchausen has videos showing exactly how to do it. Check it out.
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AMU Q&A If you don't have a plan of your own, you'll become a part of somebody else's.
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bodhisatta 
Smurf real estate agent


Registered: 04/30/13
Posts: 61,889
Loc: Milky way
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Re: Need advise on Inoculation with Agar Wedge Vs LI Vs LC [Re: Grey]
#21842220 - 06/22/15 04:19 PM (8 years, 7 months ago) |
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1. Yes 2. Irrelevant question
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MysticPop
AlienInWonderland



Registered: 05/05/15
Posts: 132
Last seen: 2 months, 30 days
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Re: Need advise on Inoculation with Agar Wedge Vs LI Vs LC [Re: bodhisatta]
#21842230 - 06/22/15 04:22 PM (8 years, 7 months ago) |
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So if i have 2 monotubs of 60 QT size.. How many Quart jars should i inoculate?
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Grey
⇜ ✯ ⇝



Registered: 11/06/14
Posts: 6,223
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Re: Need advise on Inoculation with Agar Wedge Vs LI Vs LC [Re: Grey]
#21842233 - 06/22/15 04:22 PM (8 years, 7 months ago) |
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How's the transfers looking?
If those are the plates we thought had bacteria, I wouldn't use those. Use only clean donor plates.
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AMU Q&A If you don't have a plan of your own, you'll become a part of somebody else's.
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MysticPop
AlienInWonderland



Registered: 05/05/15
Posts: 132
Last seen: 2 months, 30 days
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Re: Need advise on Inoculation with Agar Wedge Vs LI Vs LC [Re: Grey]
#21842246 - 06/22/15 04:28 PM (8 years, 7 months ago) |
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The transfer was done only yesterday.. I was planning on using these leftover plates. http://www.shroomery.org/forums/showflat.php/Number/21840040. the others would take more time i think ..What do u think about the 4 plates in the pic in the above link?
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mushpunx
Fungus Punk



Registered: 04/20/14
Posts: 13,394
Last seen: 11 days, 12 hours
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Re: Need advise on Inoculation with Agar Wedge Vs LI Vs LC [Re: bodhisatta]
#21842247 - 06/22/15 04:28 PM (8 years, 7 months ago) |
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I personally stay away from liquids. In future grows you might have a situation where an LC or LI would make your work a lot easier if you feel you have mastered the skills to pull them off. But for the beginners (and intermediates like myself) I think there are better options.
I think its best to drop wedge into master jar, and G2G.
Or, if you arent comfortable with G2G, you can divide a single plate and drop a wedge into each of the jars going to spawn a tub. But once you start handling larger volumes of spawn you will definitly want to master G2G because all wedge A2G becomes a bit of work
The key is to stagger your spawn production. Once you find a really killer culture that you like working with constantly produce spawn instead of doing one really big run. Then you will always have spawn, the few days extra it might take to colonize become immaterial.
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 Amateur Mycologists United AMU Q&A
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mushpunx
Fungus Punk



Registered: 04/20/14
Posts: 13,394
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Re: Need advise on Inoculation with Agar Wedge Vs LI Vs LC [Re: MysticPop]
#21842253 - 06/22/15 04:31 PM (8 years, 7 months ago) |
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Quote:
MysticPop said: So if i have 2 monotubs of 60 QT size.. How many Quart jars should i inoculate?
You will want 4 - 5 qts per tub. So if you are droppimg an agar wedge, you could get away with innoculating one master jar to G2G into 10 reciving jars.
If you are going all A2G you will have to inoculate 8-10 jars with wedge.
LC or LI same thing, inoculate 8-10 jars.
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 Amateur Mycologists United AMU Q&A
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MysticPop
AlienInWonderland



Registered: 05/05/15
Posts: 132
Last seen: 2 months, 30 days
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Re: Need advise on Inoculation with Agar Wedge Vs LI Vs LC [Re: mushpunx]
#21842300 - 06/22/15 04:47 PM (8 years, 7 months ago) |
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So i plan to inoculate 4-6 jars with the plates i have.. Then depending on the quality of inoculation.. I could g2g from one that is the best? I also have around 12-14 plates that are currently inoculated and resting after my A2A transfer from yesterday..
Also why is G2G considered difficult than A2G?
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mushpunx
Fungus Punk



Registered: 04/20/14
Posts: 13,394
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Re: Need advise on Inoculation with Agar Wedge Vs LI Vs LC [Re: MysticPop]
#21842546 - 06/22/15 05:53 PM (8 years, 7 months ago) |
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Quote:
MysticPop said: So i plan to inoculate 4-6 jars with the plates i have.. Then depending on the quality of inoculation.. I could g2g from one that is the best? I also have around 12-14 plates that are currently inoculated and resting after my A2A transfer from yesterday..
Also why is G2G considered difficult than A2G?
You wont likely be able to pick out which one is "best" unless some go green or something. Contams hide so well in spawn you dont usually see contamination untill you fruit your tubs.
So it would be better to either inoculate one jar, and use it to G2G into the rest of the reciving jars or just drop a wedge from the plate into each of the jars going into one tub. One or the other.
Either way make sure your plate is clean, when in doubt make another transfer or two. I usually make 3 transfers before going to grain if its from spore.
A2G is easier because you are just quickly dropping a wedge into a jar and closing it back up. G2G is tricky because of the motions.. you might accidently slip your hand over the open jar when pouring, or if you didnt wipe the jar down good enough knock contams into the reciving jar etc.. Its not that hard just a little tricky. IME most contams get into spawn during G2G so you wanna make sure your technique is spot on.
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 Amateur Mycologists United AMU Q&A
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mrbart4444
The mycelium whisperer


Registered: 09/13/14
Posts: 2,266
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Re: Need advise on Inoculation with Agar Wedge Vs LI Vs LC [Re: MysticPop]
#21842565 - 06/22/15 05:58 PM (8 years, 7 months ago) |
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I myself like to innoc a lc wih my agar and then make syringes from the growth . I dont like g2g its way easier to just inject the material
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Edited by mrbart4444 (06/22/15 05:59 PM)
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