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OfflineWhoWhatWhen
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need the pros help
    #2909807 - 07/20/04 09:08 PM (12 years, 4 months ago)

Hey all, ive been growing for probbably over a year or 2 now and im moving on to more advance tech's. so here i go.

I work at a microbiology lab so i have access to everything Hoods sterile enviroment everything. I was wondering would this work.

If i use a spore syringe to inoculate a agar dish and then put it in to a incubator tell growth apears. After the growth has become large anuff to see could i some how take cuts or "cubes" of the mycilum and then open up freshly sterile PFteks and just add it to the jar seal it up tight and Bang! let those babys grow???? would this be possible... any help would be greatly opreciated.

Thanks!


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OfflineWhoWhatWhen
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Re: need the pros help [Re: WhoWhatWhen]
    #2909812 - 07/20/04 09:09 PM (12 years, 4 months ago)

Or if any faster and greater methods are known would be cool.


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InvisibleMykey
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Re: need the pros help [Re: WhoWhatWhen]
    #2909968 - 07/20/04 09:48 PM (12 years, 4 months ago)

Sure,people have been doing it for ages. Whole grain jars would be a better recipient for those agar wedges though IMO.


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OfflineWhoWhatWhen
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Re: need the pros help [Re: Mykey]
    #2910303 - 07/20/04 11:11 PM (12 years, 4 months ago)

I've tried grain before with 1pint jars I beleive and the Filter patchs.... everytime I start them i get Black mold outbreak or some sort of mold growth. and such.... my only sucess has been with the PFTEK.... any help on making it work....


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Invisiblemycofile
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Re: need the pros help [Re: WhoWhatWhen]
    #2910662 - 07/21/04 12:41 AM (12 years, 4 months ago)

No offense, but how can you possibly work in a microbiology lab and not be able to sterilize and innoculate a jar of grain?
*edit*
sorry that wasn't helpful. At what stage do your jars of grain contaminate? Before innoculation? After? What prep methods do you use. Have you ever just left a sterilized jar alone to be sure that it doesn't contaminate (indicating your sterilization was incomplete) etc. Need this kind of info to give you tips on how to do it right, but in general I'd say just treat it like you would anything in a biolab and it should work.


--------------------
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PM me with any cultivation questions.

I just looked at my profile and realized I had a website at one point in time on geocities, it's not there anymore and I have no idea what I had on it. Anybody remember my website from several years aga? PM if so please.


Edited by mycofile (07/21/04 12:43 AM)


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OfflineMAGnum
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Re: need the pros help [Re: WhoWhatWhen]
    #2910697 - 07/21/04 12:53 AM (12 years, 4 months ago)

Dude, you can totally do that! The mycelium will grow out and colonize the whole jar from a single tiny fleck, let alone a wedge. Mycofile has a point, how did you get that job? The problem with the grain could be that you need more waqter or cooking time. Basically, you want water to be in every facet of the grain so it all sterilizes.


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Offlinehyphae
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Re: need the pros help [Re: MAGnum]
    #2914593 - 07/22/04 02:39 AM (12 years, 4 months ago)

Quote:

MAGnum said:
  Dude, you can totally do that!  The mycelium will grow out and colonize the whole jar from a single tiny fleck, let alone a wedge.  Mycofile has a point, how did you get that job?  The problem with the grain could be that you need more waqter or cooking time.  Basically, you want water to be in every facet of the grain so it all sterilizes.



Ok listen up guys Won't Work! at least with PFTek! remember the verm layer? Ya how'd you get that job bro? hey I'll take one too! :lol:


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Invisiblefishy1
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Re: need the pros help [Re: WhoWhatWhen]
    #2927388 - 07/26/04 12:28 AM (12 years, 4 months ago)

toss the whole culture in an eberbach or flask w/blender- and just make syringes from it, THEN you can use PF jars. :smile:. A single dish can produce hundreds of cc's of slurry. So much easier IMO-


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OfflineWhoWhatWhen
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Re: need the pros help [Re: WhoWhatWhen]
    #2983924 - 08/10/04 01:13 AM (12 years, 4 months ago)

ok but the problem im having is figuring out which Pre-Plates of agars we have which one would i want to use im thinking the Sab or the Maconti Which are used to grow fungus's yeast and such micros.... I know i wouldnt want to put them on a Bap plate right Sheep blood with 5%prox? or whould i use the Seaweed HIP? should i just copy the sheet of plates we have??


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Invisiblefastfred
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Re: need the pros help [Re: WhoWhatWhen]
    #2984924 - 08/10/04 07:19 AM (12 years, 4 months ago)

Use PDA, PDY, or MEA. Sab has been fairly dissapointing for a foaf. If you don't have any of those LBA will work, but not as well.

And I have no idea why you think a BAP would work... Are you sure you work in a lab and aren't just dreaming that? [edit: Sorry I just noticed that you weren't thinking of using it.]

Anyway, the method you outlined wouldn't work very well because it would take a fairly long time to colonize. Blenderize it up or use liquid culture and then inject it. That works far better than agar cubes or even multispore inoculation.

As to the person that mentioned the verm layer of PF jars... well a lot of people don't use it becasue it's a waste of verm. If you have contam problems at the jar stage then you have bigger problems than a little layer of verm can solve.

-FF


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OfflineSuntzu
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Re: need the pros help [Re: WhoWhatWhen]
    #2985071 - 08/10/04 10:17 AM (12 years, 3 months ago)

If you mean MacConkey agar [there might be 'Maconti', but I've never heard of it], it isn't even worth trying. It's specifically 'salty' to select for gram negatives [particularly gut bacteria]. Fungi don't do well on it.


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Offlinehyphae
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Re: need the pros help [Re: fastfred]
    #2985252 - 08/10/04 11:19 AM (12 years, 3 months ago)

>>As to the person that mentioned the verm layer of PF jars... well a lot of people don't use it becasue it's a waste of verm. If you have contam problems at the jar stage then you have bigger problems than a little layer of verm can solve. <<

Well #1 anyone following the PFTek IS using a verm layer (FACT), other wise they wouldn't be using it but a modified version of it :wink: The PFTek was designed for beginners with possible flaws in their sterile culture technique hence the buffer zone but reguardless of anything you are a wealth of knowledge and everyone should listen to you and all your insightful suggestions and assumptions fastfred, you know sometimes theres a big picture beyond the obvious that some of us cannot see because we don't/won't take the time to slow down and try to understand what it is we are trying to accomplish. It kinda reminds me of my daughter when she was 14, she was given the knowledge but the the reasoning and was on the I know everything kick well for the most part hell ya she was right about alot of things but couldn't give any reasoning behind it. Well she quickly learned (thank God) that there is often a bigger picture that is not evident to the inexperienced. This is said with all respects fastfred as you are bring it to the table and I respect that. By the way you got a PM! :wink:


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Invisiblefastfred
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Re: need the pros help [Re: hyphae]
    #2995938 - 08/12/04 04:10 PM (12 years, 3 months ago)

Quote:

dcyans said:
Well #1 anyone following the PFTek IS using a verm layer (FACT), other wise they wouldn't be using it but a modified version of it :wink:




Criticism of semantics noted.

Quote:

The PFTek was designed for beginners with possible flaws in their sterile culture technique hence the buffer zone but reguardless of anything you are a wealth of knowledge and everyone should listen to you and all your insightful suggestions and assumptions fastfred[...]




I understand your point and the reasoning behind using the verm layer.  However, I dispute the utility of it.  Any contams are most likely already in the syringe or on the needle.  The verm layer does nothing to protect against these contams since they get washed down with the innoculate.  The only way that the verm layer would protect you is against a spore falling down through the tiny injection hole.  I find that highly unlikely to be a major source of contams.  The verm layer also usually ends up being fairly moist, so contams probably would end up germinating on it anyway.


Quote:

[...]there is often a bigger picture that is not evident to the inexperienced.




The verm layer only makes sense if:
#1  It actually works
#2  People have such poor aseptic technique as to actually need it

I don't think that either of these are good assumptions to make.  I would be interested to see if anyone has compared jars with and without a verm layer.  I think there would be little, if any, difference.

Quote:

This is said with all respects fastfred as you are bring it to the table and I respect that. By the way you got a PM! :wink:




I perfer to have my errors corrected rather than defend them in perpetuity.  A slightly pompus attitude results in many people eager to correct me.

I also do have some education and experience in the subject, so I'm not just talking out my ass.

-FF


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Offlinehyphae
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Re: need the pros help [Re: fastfred]
    #2997032 - 08/12/04 08:21 PM (12 years, 3 months ago)

I grew up with PF's tek and have seen it make indoor growing boom so in part i'm defending it FF. I can see the verm layer as working in several ways which are unique to this tek only. #1 is that these cakes were meant to be turned upside down dislodging the cake loose with a sharp rap, and then the verm layer stays at the base in the cover and acts as a moisture reservoir and is usually where the shrooms like to really pin. #2 with the 4 holes as suggested and the verm layer makes for excellent gas exchange and no tyvek/hepa/coffee filter needed. #3 as suggested the needle needs to be flamed or wiped in alcohol and any possible contams left would be wiped back as the needle is inserted thru the verm layer. These may be all be moot points to you as you are experienced but this tek was not meant for you or me. BTW I notice immediately you weren't a newbie bro! :wink: take care FF


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Invisiblefastfred
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Re: need the pros help [Re: hyphae]
    #2998639 - 08/13/04 02:18 AM (12 years, 3 months ago)

Instead of the verm you can add more substrate, producing better yeilds IMHO.

They still pin great from the bottom without the verm layer. If you let the mycelium come directly into contact with perlite it can really suck up a lot of water that way.

PF's Tek isn't just for beginners, IMHO. While casing works wonders and gives the best yeild, PF lets you run a bunch of experiments in a small area under identical conditions.

Just my 2 cents... Keep it real dcyans.


-FF


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OfflineRaadt
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Re: need the pros help [Re: fastfred]
    #2999141 - 08/13/04 05:33 AM (12 years, 3 months ago)

try making a grain jar, and leaving just a pinhole.

I don't think you'll have much success.

The verm barrier is a non-nutritive layer, for any 'filth' to become trapped, before hitting the nutritive substrate.

Use some common sense. Make 10 pf jars, with no verm layer, and leave them uncovered in an incubator. Make 10 more with a verm layer, and do the same.

What do you want to bet contamination is higher in the one without?

A pinhole is like a fucking empire state building sized tunnel for bacteria.


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Offlinescatmanrav
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Re: need the pros help [Re: Raadt]
    #2999331 - 08/13/04 08:26 AM (12 years, 3 months ago)

I think his point is, neither would be successful if left uncovered. Dont you usually wait until the jar is partially colonized giving it a better chance to fight off molds? It seems whenever people remove the foil to early I see them posting they got contams. I'm talking mostly out of my ass here as I've never done the PF Tek (though reviewed it fully and in variations) but thats just what Ive seen people post about.

If I was to use the PF Tek, which I'm sure I will some day when I feel ambitious, I wouldnt use the verm layer, as fastfred said..more substrate per jar. Though I would use Tyvek and PC em, then why would you really need it? I mean shouldnt be any different then grains but you dont shake it and knock in 4 places.


--------------------
"life is like a drop of rain getting closer and closer to falling into a lake, and then when you hit the lake there is no more rain drop, only the lake."

Growing with bags, start to finish (including my new grain and substrate prep)
Anyone looking to start bulk tubs/mono tubs/shotgun hybrids? Good tubs to use..
How I do grain (old still good tips)
Turn your closet into a fruiting chamber
Casing layer colonization and overlay


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Offlinehyphae
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Re: need the pros help [Re: fastfred]
    #2999366 - 08/13/04 08:45 AM (12 years, 3 months ago)

Quote:

fastfred said:
Instead of the verm you can add more substrate, producing better yeilds IMHO. 

They still pin great from the bottom without the verm layer.  If you let the mycelium come directly into contact with perlite it can really suck up a lot of water that way.

PF's Tek isn't just for beginners, IMHO.  While casing works wonders and gives the best yeild, PF lets you run a bunch of experiments in a small area under identical conditions.

Just my 2 cents...  Keep it real dcyans.


-FF


FF Leave the cakes in direct contact with the perlite? thats been a nono for years (clue) contam city and can also lead to oversaturation of the cakes. ALSO have ya ever heard of double ending cakes? this actually leads to better yields. Your (nothing personal) kinda talking like a newbie but keep on this is getting very enlighting. Ya know I've been around for years and have stayed away from the Shroomery bc of this sort of stuff and decided to mod at a sister site instead but the draw of teaching has brought me back. I'm not here to argue FF now I've done many a vermless cake when doing the lazymans ziplock tek for expanding substrate so I known all about vermless cakes it's also been around for years :wink: GF FF
And scatmanrav you could leave a PFTek cake uncovered and the substrate would be perfectly safe from contams FYI.


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OfflineGr0wer
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Re: need the pros help [Re: hyphae]
    #2999549 - 08/13/04 09:56 AM (12 years, 3 months ago)

I must say cakes dont seem to be as you are saywing with consistancy for experimenting. Most BRF grows i see have uneven growth with 1-3 excelling cakes for no reason. You need a larger mass to becomre more stable with experiments, this is where bulk steps in. Most people dont seem to be doing crazy experiments besides just having the ideal conditons and growing perfect cubes.


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Invisiblefastfred
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Re: need the pros help [Re: Raadt]
    #3000605 - 08/13/04 02:11 PM (12 years, 3 months ago)

Quote:

Raadt said:
try making a grain jar, and leaving just a pinhole.

I don't think you'll have much success.




I've never even heard of using a verm layer with grain! There is a inoculation hole at the top of every grain jar, and I have great success with them ( < 1-2% contam rate).


Quote:

The verm barrier is a non-nutritive layer, for any 'filth' to become trapped, before hitting the nutritive substrate.




It's far easier to simply omit the "filth" from the jars.


Quote:

Use some common sense. Make 10 pf jars, with no verm layer, and leave them uncovered in an incubator. Make 10 more with a verm layer, and do the same.




It's been done... and I saw no difference. That doesn't mean that it isn't helpful for those that can't use a bunsen burner, or build a flow hood, or use a glove box. But, frankly, who are those people? I would imagine mostly kids, hiding this stuff under their beds. They shouldn't be growing anyway, and I think helping them is NOT the right thing to be doing IMHO.


Quote:

What do you want to bet contamination is higher in the one without?




I've never done a sizeable enough load with verm layer cakes to make the 1-2% contam rate yield a statistically significant result. At those levels, I would have to do several hundred verm layer jars just to get a general idea of whether there is any true value to the verm layer. By that time the increased amount of substrate in the non-verm layer jars would have easily negated the 2-4 contams that I would likely see. That even assumes that the verm layer works 100% of the time. If it only works 50% of the time then it would take 200 jars to see a reduction of comtamed jars by 2... Hardly worth it.

If OTOH, you are getting a 20% contam rate then you might see a benefit. If you are getting that many contams though, you have more problems than a simple layer of verm will prevent IMHO. If you can cut your contam rate from 20% to 10%, this is still unacceptable to me, and I think you will get more mileage out of improving your aseptic technique.


Quote:

A pinhole is like a fucking empire state building sized tunnel for bacteria.




Spores fall through the air very slowly. You should only have the hole exposed for 5 sec. or so, depending on how many holes you use. Even in open air, without a bunsen burner, this shouldn't be much of a problem for you.


-FF


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Mushrooms, Mycology and Psychedelics >> Advanced Mycology

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