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Damion5050
Mush Doctor


Registered: 05/01/08
Posts: 12,493
Loc: Lost In Translation !
Last seen: 3 years, 11 months
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Re: cloning question [Re: hamloaf]
#13013614 - 08/06/10 05:21 PM (14 years, 5 months ago) |
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I mainly use LC's when doing like spawn bags or if I need to knock up a bunch of jars in a hurry.. Other wise I take my time and do agar to grain the g2g
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hamloaf
Q-dood ®©™√


Registered: 12/23/09
Posts: 24,389
Loc: ation: Based.
Last seen: 1 day, 2 hours
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Re: cloning question [Re: Numinosum]
#13013632 - 08/06/10 05:25 PM (14 years, 5 months ago) |
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Quote:
Numinosum said: Oh, btw HL, I just bought a 40lb of rice bran to try out. I will post the results in your thread.

Cool. Do that. You won't be disappointed that you replaced BRF for Rice Bran, I guarantee it!. For cubes, I will never use BRF again. How did you like that price?
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Numinosum
President of Turd Town



Registered: 05/19/09
Posts: 1,175
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Re: cloning question [Re: hamloaf]
#13013647 - 08/06/10 05:28 PM (14 years, 5 months ago) |
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Yeah 40lbs for $22. I don't usually use brf but after reading your thread I got motivated to go get some rice bran and make some blocks.
-------------------- ...within my memory is the knowledge of hyper-light drive ships and how to build them.
Doc_T's Efficiency Challenge
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hamloaf
Q-dood ®©™√


Registered: 12/23/09
Posts: 24,389
Loc: ation: Based.
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Quote:
Damion5050 said: I mainly use LC's when doing like spawn bags or if I need to knock up a bunch of jars in a hurry.. Other wise I take my time and do agar to grain the g2g
Me too, exactly except I don't get in a hurry. LC's to spawn bags. I knock my jars up with agar always, unless I am testing LC. I am currently breaking that habit of LC and am going for the agar wedge to spawn bag and g2g with spawn bags in front of a flowhood. I have had it with LC's, agar is soooo much better, cheaper, efficient, concise, and easier than LC. Plus it makes you feel like a mad scientist. HL
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Damion5050
Mush Doctor


Registered: 05/01/08
Posts: 12,493
Loc: Lost In Translation !
Last seen: 3 years, 11 months
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Re: cloning question [Re: hamloaf]
#13013673 - 08/06/10 05:33 PM (14 years, 5 months ago) |
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LoL the only down side to agar in some cases is the amount of wedges u need to use to get a quart jar to colonize in a timely manner.
I sometimes use like a 1/2 pint jar the do grain LC with that to speed it up some.
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Numinosum
President of Turd Town



Registered: 05/19/09
Posts: 1,175
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I'll be making my first inoculations with agar tonight, I figure I'll take a tiny piece to a new dish, then use the rest for my jars, bags. I have a different "substrain" to test on each.
-------------------- ...within my memory is the knowledge of hyper-light drive ships and how to build them.
Doc_T's Efficiency Challenge
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hamloaf
Q-dood ®©™√


Registered: 12/23/09
Posts: 24,389
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Quote:
the only down side to agar in some cases is the amount of wedges u need to use to get a quart jar to colonize in a timely manner.
When using single section petri dishes or pint jars for agar to make MS wedges for fast colonization, I cut my agar into wedges like a pizza. I get 8 wedges out of a jar or single section petri dish and never have any colonization issues as far as speed is concerned and I don't shake. I think I can do the same with half the amount of an agar wedge too, getting 16 wedges out of a single section petri of or a jar. It truly is amazing, how in this particular hobby, like no other, the things that work for one doesn't work for another. HL
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wygram
Myconaut

Registered: 01/28/07
Posts: 573
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Re: cloning question [Re: hamloaf]
#13013890 - 08/06/10 06:16 PM (14 years, 5 months ago) |
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Quote:
hamloaf said: [I get 8 wedges out of a jar or single section petri dish and never have any colonization issues as far as speed is concerned and I don't shake.
Those are some giant wedges. You only really need a tiny rice sized piece, then once the mycelium takes to the grain to about a quarter sized growth shake to redistribute and then shake once again at the normal 30%.
-------------------- Changing your mind is one of the best ways of finding out whether or not you still have one.
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hamloaf
Q-dood ®©™√


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Re: cloning question [Re: wygram]
#13014056 - 08/06/10 06:54 PM (14 years, 5 months ago) |
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Quote:
wygram said: Those are some giant wedges. You only really need a tiny rice sized piece, then once the mycelium takes to the grain to about a quarter sized growth shake to redistribute and then shake once again at the normal 30%.
Yes wygram, they are. I have become aware of that the agar wedges I was using were excessive, through trial and error. I don't shake my jars at all, not once during colonization, as stated earlier, I am never in a rush (when it comes to my grows). It barley happens but I just hate it when my jars don't recover from the shake. It get's me fightin' mad. I knead apart spawn bags however, at 25 percent colonization because spawn bags allow you to be more gentle with the myc come time to "shake". I just drop a wedge onto the top of the grains and two weeks later, full colonization. It was nice to talkin' to you, wygram. Have a good weekend and please, POAST MOAR!!
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RogerRabbit
Bans for Pleasure


Registered: 03/26/03
Posts: 42,214
Loc: Seattle
Last seen: 1 year, 10 months
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Quote:
OMackaRoni said: it's intimidating to me because of the use of agar and a petri dish. those things sound very high tek to my fairly noob mind.
Watch the agar/petri dish video, the cloning video, and the strain isolation video. Bear in mind, these are only sample clips from the DVD, but they'll get the point across and it won't seem so hard. Once you get those down, watch the master culture slant video to see how we store our cultures for long term use. RR
-------------------- Download Let's Grow Mushrooms
semper in excretia sumus solim profundum variat
"I've never had a failed experiment. I've only discovered 10,000 methods which do not work."
Thomas Edison
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Javadog
Continuing along



Registered: 05/03/10
Posts: 7,385
Loc: USA
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Quote:
Damion5050 said: I think the same way I don't know why more people don't use agar... I mean it really aint that hard. After my second try or so with agar I had my technique down and knew what I was doing and now it is super super easy..
While I had planned to move toward agar, and have already made a batch and poured my first dishes...I must admit that I am being forced to advance the schedule as I am having trouble with infected LCs.
This is good in a way, as I will be working with prints, and some I made myself, so agar was coming.
-------------------- Boyd Rice told my brother that life is a corny pack of freesakes
Myco-tek.org
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wygram
Myconaut

Registered: 01/28/07
Posts: 573
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Re: cloning question [Re: Javadog]
#13015309 - 08/07/10 12:09 AM (14 years, 5 months ago) |
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Quote:
Javadog said: This is good in a way.
Yes it is. Agar is the superior spore starting method because you can ensure you have a clean culture before inoculating another medium. Since mushrooms are grown under non-sterile conditions, there is always a chance a print will contain some contaminant spores.
And it really is...
Quote:
Damion5050 said: super super easy..
-------------------- Changing your mind is one of the best ways of finding out whether or not you still have one.
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anathema
pǝsnɟuoƆ



Registered: 03/13/05
Posts: 44
Loc: BC, Canada
Last seen: 12 years, 3 months
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Re: cloning question [Re: wygram]
#13015404 - 08/07/10 12:31 AM (14 years, 5 months ago) |
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I know it is more of an agar thread as opposed to LC but I really am enjoying the mycellium water method.
agar (the poster) posted this tek:
http://www.shroomery.org/forums/showflat.php/Cat/0/Number/6817701/an/0/page/0
You start with a healthy grain jar so are almost certain you have a clean starting point, since it is usually easy to tell if you have a contam'd grain jar.
I injected my grain jar with water, sucked it out into a few 20cc syringes, then later used the myc water, worked perfectly and much faster than multispore.
Being almost sure of a clean sample, plus fast colonization etc is great. No way to isolate as per agar of course, but thats life.
-------------------- --
Not all those who wander are lost.
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Javadog
Continuing along



Registered: 05/03/10
Posts: 7,385
Loc: USA
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Re: cloning question [Re: wygram]
#13015439 - 08/07/10 12:39 AM (14 years, 5 months ago) |
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Quote:
wygram said:
Quote:
Javadog said: This is good in a way.
Yes it is. Agar is the superior spore starting method because you can ensure you have a clean culture before inoculating another medium. Since mushrooms are grown under non-sterile conditions, there is always a chance a print will contain some contaminant spores.
And it really is...
Quote:
Damion5050 said: super super easy..
The agar dishes did confirm that my syringes are contaminated with bacteria. I want to do the hot pour method rather than try to take a clean sample.
Do you recall the temperature that this should be done at?
Actually I posted this in a separate thread, with photos, so I will stop here.
-------------------- Boyd Rice told my brother that life is a corny pack of freesakes
Myco-tek.org
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hamloaf
Q-dood ®©™√


Registered: 12/23/09
Posts: 24,389
Loc: ation: Based.
Last seen: 1 day, 2 hours
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Re: cloning question [Re: Javadog]
#13015969 - 08/07/10 06:08 AM (14 years, 5 months ago) |
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Quote:
The agar dishes did confirm that my syringes are contaminated with bacteria.
What makes you so sure that the agar confirmed that your syringe(s) are contaminated? How do you know it's not your sterile procedure?
Quote:
I want to do the hot pour method rather than try to take a clean sample. Do you recall the temperature that this should be done at?
Not specific temperature is applied. Pour your agar once the container has cooled just enough to be handled with your hands but still quite hot. Just typing this reply and thinking about pouring agar is making my hands burn. HL
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Javadog
Continuing along



Registered: 05/03/10
Posts: 7,385
Loc: USA
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Re: cloning question [Re: hamloaf]
#13017892 - 08/07/10 04:02 PM (14 years, 5 months ago) |
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Ham,
Well, what I have to go on is my limited experience.
I had been making one good LC after another, posting comments like the common "Karo < SITR < Honey < LDM+Dex"....and then I started these next four strains and every LC batch of the three syringes went bad.
As to technique I place the jars in front of my flow hood to cool and work there once they are cooled. I flame sterilized the needle before each jar.
So, while I may have to admit there are other possibilities, when I drop some spore solution onto agar dishes and see all three of the problem strains show bacterial contams, where the one strain that made clean LC is not, then I am inclined to suspect the source of the spore solution.
I will work to clean these up on agar. I need to confirm the temperature that the hot agar is poured over the infected plate.
I am making ten new dishes today, and will have something to transfer to once they grow through to the surface.
I just cannot allow Cambodian, Tanseki, and Elephant Dung slip away!!!
JD
-------------------- Boyd Rice told my brother that life is a corny pack of freesakes
Myco-tek.org
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