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OfflineMighty Bop
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What Do You Think I Did Wrong?
    #1628382 - 06/12/03 10:31 AM (13 years, 5 months ago)

OK the first two times I grew, everything was a success. I made some prints and didn't try again for almost a year later.
Anyways, I'm pretty sure I did everything in the procedure correctly, so I don't think it's that. But let me tell you everything I did and then what I think may have been the problem. I'd also like others to comment as well. Maybe there is something I missed.

1. I boiled a new syringe in some distilled water, sucking in and squirting out the water. Finally I filled the syringe with water and let it sit overnight.

2. Turning on the oven to low heat and pulling out one of the shelves, I took my clean shot glass, my print and my syringe and set everything up on the shelve to make the syringe. I was wearing a surgical filter mask as well. I scraped the spores into the shot glass. Then I squirted water into the shot glass to mix around the spores. Finally I sucked the spores into the syringe one last time and I was done. I let the syringe sit for at least 24 hours.

3. I mixed all the right ingredients in the right ratio for my substrate, which was BRF/verm with distilled water.

4. I covered the wide-mouth jars with some filters then sterilized in my pressure cooker for one hour. All the jars were left to cool overnight.

5. I took all my jars and my syringe and set up on the oven shelf again.

6. I began innoculating the jars with 1CC each. I wiped the needle with alcohol after each jar.

7. After all jars were innoculated I let them sit in a shoebox in the dark with a temp of around 75 degrees.

8. After around 5 days the green mold begins.

So that's it. I think my procedure and sterilty precautions are fine because I did it the exact same way the very first two times I grew.

Now here are the variables that I think could have contributed to the failure:

1. The spores were too old.
2. The vermiculite I used was different and absorbed too much water.
3. I let all the steam out of the pressure cooker without letting it relieve the pressure by itself.

Those are basically the only things I did diferently. I am thinking it was the spores being too old, but I don't know. I thought spores last a really long time. I guess when I made the prints something went wrong. (Although the second attempt at growing was a success and I used a self-made print.)

Anyways, any comments are appreciated. Thanks!



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"One attaineth whatever state of being one thinketh about at the last when relinquishing the body, being ever absorbed in the thought thereof." - Sri Krishna to Arjuna, Bhagavad Gita, viii, 6


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OfflineFuman
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Re: What Do You Think I Did Wrong? [Re: Mighty Bop]
    #1628391 - 06/12/03 10:38 AM (13 years, 5 months ago)

Quote:


2. Turning on the oven to low heat and pulling out one of the shelves, I took my clean shot glass, my print and my syringe and set everything up on the shelve to make the syringe. I was wearing a surgical filter mask as well. I scraped the spores into the shot glass. Then I squirted water into the shot glass to mix around the spores. Finally I sucked the spores into the syringe one last time and I was done. I let the syringe sit for at least 24 hours.




I would bet the contams came during this stage. It sounds like you took some good precautions, but I am not sure that just using the oven is enough. A healthy spray of Lysol in a room with all fans turned off may help. Recently there was a post about grain to grain that talked about using a black plastic bag when transfering grain. Something like that may help.
Keep Trying


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OfflineMighty Bop
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Re: What Do You Think I Did Wrong? [Re: Fuman]
    #1630129 - 06/13/03 12:16 AM (13 years, 5 months ago)

But the thing is, the first two times I did it the exact same way so I don't think this was the problem.


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"One attaineth whatever state of being one thinketh about at the last when relinquishing the body, being ever absorbed in the thought thereof." - Sri Krishna to Arjuna, Bhagavad Gita, viii, 6


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Offlinegourmetbog
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Re: What Do You Think I Did Wrong? [Re: Mighty Bop]
    #1630229 - 06/13/03 01:12 AM (13 years, 5 months ago)

Also make sure your oven is super clean. A dirty oven is gonna have a heap of nasty bacteria. Give your oven a good clean.


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Invisiblemicro
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Re: What Do You Think I Did Wrong? [Re: gourmetbog]
    #1630318 - 06/13/03 02:00 AM (13 years, 5 months ago)

2 times might have been luck; the oven tek is said not to work very well. Could also be the spores, though, if the foreign spores got in at this point.

--
Micro


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OfflineExtravagantDream
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Re: What Do You Think I Did Wrong? [Re: micro]
    #1630344 - 06/13/03 02:11 AM (13 years, 5 months ago)

I agree that your failure came from making the syringe. It is not necessarily that you did anything wrong.. maybe just some bad luck.. who knows maybe something fell out of your hair. It is really hard to say, but it happens to everyone.

One other thing that I would like to suggest is to PC your syringes for 30 min. I had some recent failures with repeated bacterial contamination until it was suggested to me that they came from the syringe and that it should be PCed. Good luck.


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OfflineChrisH
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Re: What Do You Think I Did Wrong? [Re: ExtravagantDream]
    #1630386 - 06/13/03 02:29 AM (13 years, 5 months ago)

Mabey your spore print was contaminated?


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OfflineMighty Bop
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This Is Ridiculous [Re: ChrisH]
    #1764497 - 07/31/03 12:57 AM (13 years, 4 months ago)

OK so I got some more spores and we tried again. Used face masks, gloves, the oven tek, alcohol wipes and flame sterilizing the needle. In other words, we did everything right. BUT, there were some things we did do a little different, to save time, just like last time. First we manually let the steam out of the pressure cooker without letting it depressurize itself. Then all the jars were placed in the fridge to let cool for a couple hours. Then they were set out at room temperature for an hour before we innoculated. We did everything right but we still got green and black and spiderweb mold on most of the jars. What the hell are we doing wrong??

I have narrowed it down to a number of things we did differently, but really don't make much sense as to why it would matter. And some of them are the same things as my first post, almost two months ago. They are:

1. Using a different kind of vermiculite than my first two grows.
2. Letting the steam out manually from the pressure cooker.
3. Not letting the jars cool overnight.

I have no idea why any of these things would really matter that much, but these are the only factors that are different. I think I am gonna try grain the next time, which will take the vermiculite factor out of the equation. What do you guys think?


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"One attaineth whatever state of being one thinketh about at the last when relinquishing the body, being ever absorbed in the thought thereof." - Sri Krishna to Arjuna, Bhagavad Gita, viii, 6


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Offlinedickmcplentie
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Re: This Is Ridiculous [Re: Mighty Bop]
    #1764544 - 07/31/03 01:12 AM (13 years, 4 months ago)

I could be wrong but I doubt that the 3 reasons above have anything to do with it.

1. If the verm is PC'ed it wont contaminate
2. I have done this with wonderful results and it shouldn't make any difference at all
3. cool is cool, yes the rapid cooling can "suck" in contams but if you seal your jars right this isnt an issue

The fact that you seem to loose 100% of your jars is interesting. You have a contaminated source or your doing something wrong and don't know it.

Did the new spores come from your first two grows?
How long do you PC and at what PSI?
How do you seal/filter your jars during PC'ing?
Do you use pond water? just kidding

If it does turn out to be contaminated prints try to remember that prehaps not all the prints were contaminated. That may explain why you had the second success. Or maybe the prints got contaminated in the year of storage.
Or you could be suffering from my same problem and its just that you have fucked your kharma up so bad you are going to have to work twice as hard at everything you do.


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The above statement has been a figment of my imagination. Niether the statement nor the writer is real, we are both fiction & should not be taken seriously in any way


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Offlinewhole9
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Re: This Is Ridiculous [Re: dickmcplentie]
    #1764718 - 07/31/03 02:23 AM (13 years, 4 months ago)

Try not using jars with filters. You really dont need them. Just use tape to cover the holes.


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OfflineMighty Bop
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Re: This Is Ridiculous [Re: dickmcplentie]
    #1765346 - 07/31/03 09:26 AM (13 years, 4 months ago)

I got the spores from TLG, so I'm sure they were good. I PC'ed the jars for 1 hour each. Maybe I should have done it longer? I have one of those cheap little PC's and I cannot tell the temp. I used Micron Magick filters on the jars. And I used distilled water. I just can't figure it out. It makes no sense to me. Also, with my first successes, I used an incubation chamber. I doubt this would matter very much, though.

For some reason I think the vermiculite is to blame. I don't know why though. Maybe this new kind needs longer to PC? I think the first time I used Scott's brand or Schulz but I can't find these anywhere. I think they took them off the market. So I had to use this third party brand verm. Any more input guys?


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"One attaineth whatever state of being one thinketh about at the last when relinquishing the body, being ever absorbed in the thought thereof." - Sri Krishna to Arjuna, Bhagavad Gita, viii, 6


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Offlinescreaminsemen
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Re: This Is Ridiculous [Re: Mighty Bop]
    #1765378 - 07/31/03 09:39 AM (13 years, 4 months ago)

Stupid question, but did you cover your jars during pcing with foil? And maybe raising the incubation temp to 86 degrees.


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Offlinedickmcplentie
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Re: This Is Ridiculous [Re: screaminsemen]
    #1765512 - 07/31/03 10:58 AM (13 years, 4 months ago)

Sounds like you are doing everything right. If you suspect the verm go buy some more, its cheap. I REALLY don't think the brand matters at all but perhaps you got a nasty bag *lifts eyebrows

maybe you arent getting to 15 PSI on the PC?
Perhaps its time to hit a thrift store for a $10 PC that has a gauge?

The 100% lose is telling. Try not innocing one jar next test. Do all the others like you would normally would but leave one sealed after PC'ing. If you loose all but this one it could be the spores. If you loose this one too its the substrate. If you don't loose any...
inoc>spawn>case>fruit>enjoy


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The above statement has been a figment of my imagination. Niether the statement nor the writer is real, we are both fiction & should not be taken seriously in any way


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OfflineMighty Bop
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Re: This Is Ridiculous [Re: dickmcplentie]
    #1766757 - 07/31/03 06:20 PM (13 years, 4 months ago)

As a matter of fact, I DIDN'T cover my jars with tin foil while PC'ing. And I think the first two times it worked I did. Most of the filters had gotten water on them after I was done cooking. Could this cause problems? If so, this could be the answer.

And I'm not sure if the pressure is getting up to 15PSI, there is no gauge. I agree, I need to get a canner. Do they have these at thrift stores?


--------------------
"One attaineth whatever state of being one thinketh about at the last when relinquishing the body, being ever absorbed in the thought thereof." - Sri Krishna to Arjuna, Bhagavad Gita, viii, 6


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Offlinemr_minds_eye
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Re: What Do You Think I Did Wrong? [Re: Mighty Bop]
    #1766839 - 07/31/03 06:44 PM (13 years, 4 months ago)

I've always used the oven tek and it hasn't let me down.


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Our quest for discovery fuels our creativity in all fields, not just science. If we reached the end of the line, the human spirit would shrivel and die. But I don't think we will ever stand still: we shall increase in complexity, if not in depth, and shall always be the center on an expanding horizon of possibilities.
-Stephen Hawking


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OfflineMighty Bop
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Re: This Is Ridiculous [Re: screaminsemen]
    #1766859 - 07/31/03 06:49 PM (13 years, 4 months ago)

I'm beginning to think more and more that not using the tin foil while PC'ing was the problem, but why??


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"One attaineth whatever state of being one thinketh about at the last when relinquishing the body, being ever absorbed in the thought thereof." - Sri Krishna to Arjuna, Bhagavad Gita, viii, 6


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OfflineSev
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Re: This Is Ridiculous [Re: Mighty Bop]
    #1767423 - 07/31/03 10:45 PM (13 years, 4 months ago)

Quote:

mighty_bop said:
As a matter of fact, I DIDN'T cover my jars with tin foil while PC'ing. And I think the first two times it worked I did. Most of the filters had gotten water on them after I was done cooking. Could this cause problems? If so, this could be the answer.





It's not a problem.  My tyvek filters always have water on them -- I just shake it off when I take the lid off to innoc. 

Quote:


And I'm not sure if the pressure is getting up to 15PSI, there is no gauge. I agree, I need to get a canner. Do they have these at thrift stores? 




Canners are expensive, and not -really- worth it if you're just starting out and already have a PC.  If your'e doing a lot of jars or quart stuff, on the other hand...but, anyway.

It really sounds to me like your syringe was contamn'd, or the spore print you made it from was.  Even if you were just boiling, you shouldn't've gotten as high a fail rate as you did.  Gotta work on your syringe tek.  :smile:


--------------------
"Do we want the stars? We can have them. Can we borrow cups of fire from the sun? We can and must and light the world." --"On the Shoulders of Giants", Ray Bradbury

All of my posts are full of fiction and blatant lies.


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OfflineMighty Bop
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Re: This Is Ridiculous [Re: Sev]
    #1768402 - 08/01/03 05:58 AM (13 years, 4 months ago)

I got my syringes from TLG actually and I'm pretty sure they were viable, but who knows. I am thinking that the first kind of verm I got was pre-sterilized, meaning I didn't have to PC it for that long, and this new stuff is not pre-sterilized. I don't know. I guess I just have to keep changing the variables around. It sucks because it's getting expensive. But I will grow again!


--------------------
"One attaineth whatever state of being one thinketh about at the last when relinquishing the body, being ever absorbed in the thought thereof." - Sri Krishna to Arjuna, Bhagavad Gita, viii, 6


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OfflineMighty Bop
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Re: This Is Ridiculous [Re: Mighty Bop]
    #1897451 - 09/09/03 09:55 AM (13 years, 2 months ago)

Well what do you know, we tried again and it failed. I have NO IDEA what we're doing wrong, since we are basically doing everything by the book. My only guesses as to what went wrong are the following......

1. somehow our BRF and verm are no good
2. we are not sterilizing long enough (we are doing about one hour with a 6-quart PC)
3. water is getting on our micron magick filters, therefore soaking into the dry layer of verm on top, somehow contaminating the substrate
4. our spores are bad
5. we are getting contamination during innoculation

Like I said in my first post, the first two times I grew, everything went fine, which is why this is so damn frustrating. I wish there was a grow video or something showing step by step instructions or something. Anyone have iny input? Thanks.....

Mighty_bop


--------------------
"One attaineth whatever state of being one thinketh about at the last when relinquishing the body, being ever absorbed in the thought thereof." - Sri Krishna to Arjuna, Bhagavad Gita, viii, 6


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OfflineMighty Bop
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Re: This Is Ridiculous [Re: Mighty Bop]
    #1897814 - 09/09/03 12:59 PM (13 years, 2 months ago)

One thing I am gonna try differently next time is using the actual lids of the jars and punching holes in them. This should prevent water from getting into the jars, like I think they were with the filters. Plus, I am going to use a glove box. Any other suggestions?


--------------------
"One attaineth whatever state of being one thinketh about at the last when relinquishing the body, being ever absorbed in the thought thereof." - Sri Krishna to Arjuna, Bhagavad Gita, viii, 6


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