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Offlinemagickpsychonaut
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Giving up... for a while
    #14237382 - 04/04/11 05:28 PM (12 years, 9 months ago)

Ok after months and months and 9 different attempts I am giving up on cultivating. I have tried my best, spent a total of almost $1200 an entire year, and still have not been successful. I have watched RR's tech 11 times over again, read through numerous teks several times over again and followed meticulously along only to fail. I just can't figure out what I am doing wrong so I am going to take a few months break. In the mean time do you have any recommendations for me? I am open to anything, even if you think that I probably already know it please don't hesitate to give me feedback.
Thank you


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Offlineagonzalez64
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Re: Giving up... for a while [Re: magickpsychonaut]
    #14237402 - 04/04/11 05:34 PM (12 years, 9 months ago)

i think am unsucessfull 3 times, beginner also, but 9 times??, could it be the water your using? what tek are you using??


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Offlineredcat
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Re: Giving up... for a while [Re: agonzalez64]
    #14237418 - 04/04/11 05:39 PM (12 years, 9 months ago)

Other than posting in this forum you don"t say what's been going wrong.


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Offlinemagickpsychonaut
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Re: Giving up... for a while [Re: agonzalez64]
    #14237472 - 04/04/11 05:54 PM (12 years, 9 months ago)

I am using RR's Tek, but I am using a PC instead of regular boiling. Here are the steps:

1. Create substrate jars, always 5 , 2 cups vermiculite, 1 cup organic brown rice flour, 1 cup distilled water. I always have a blank jar and they always turned out clean no contams.
2. Pressure cook on 2 (15psi) for 90 minutes, cool down 1 day.
3. Clean out entire bathroom. All items including shower curtain, bathroom personals totally cleaned out. I use bleach to clean all walls, window, counters, toilet, and bathtub/shower. I also clean the walls of the bathroom.
4. Next, I shower and put on clean clothes.
5. I wipe all counters again with bleach and get out glove box. I clean the glove box with bleach and spray inside with lysol.
6. I bring the PC into the bathroom unopened.
7. I spray the bathroom with lysol and close the door. I wait 30 minutes for it to settle and then go back in.
8. I put on sterile gloves and disinfect with rubbing alcohol and hand sanitizer. Next I wipe the lighter and the syringe.
9. I take jars out and put into glove box.
10. I flame tip the needle until red hot, and wipe down with rubbing alcohol pad and take the aluminum foil off the jars and inoculate.
I flame tip after each jar.

I have done a variation of the above by adding tyvek filters, and still did not get any success.

At first I thought that the air in the bathroom may be too humid, but when I did some jars without spraying bleach water or lysol, they still turned out contaminated.


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OfflineMycelio
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Re: Giving up... for a while [Re: magickpsychonaut]
    #14237770 - 04/04/11 06:40 PM (12 years, 9 months ago)

Then the reason for failure is in your spore syringe. Contact your vendor and ask for a replacement. If it is who I have in mind, he will be happy to do so. With a clean spore syringe, you can skip 3 to 5 and 7.

Carsten


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Offlinemagickpsychonaut
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Re: Giving up... for a while [Re: Mycelio]
    #14237955 - 04/04/11 07:15 PM (12 years, 9 months ago)

I thought that it may have been the syringes, but they come from a reputable vendor and I emailed the vendor who is a highly shroomery recommended vendor who has a very good reputation. I emailed him and  he told me his stuff was good and just said too bad. (I emailed him past the 30 or 90 day guarantee) So I thought it may be something on my part. I'll order from another vendor and see if my results are different.

Also, could it be that maybe the showering before hand caused too much humidity in the bathroom where I inoculated? Or could it be that I sprayed too much lysol /bleach water? I tend to spray vigorously, because I am paranoid of getting contams.

Anyway in the mean time what I'm doing is making pf cakes and inoculated with a dummy syringe that has sterilized water in it and going through the motions  and keeping meticulous records of exactly every step, so I can track down my method and see if there are any contams. I'll be making 5 pf jars according to RR's video. 3 will be inoculated with the dummy syringe in the glove box, while one will only have the foil removed inside the glove box and put back on. The last one will serve as a blank.

Thoughts,  feedback?


Edited by magickpsychonaut (04/04/11 07:16 PM)


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OfflineSeniorCoolo
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Re: Giving up... for a while [Re: magickpsychonaut]
    #14238110 - 04/04/11 07:46 PM (12 years, 9 months ago)

You said you wipe your syringe after you flame it. Don't do that. When it's red hot it can't get any more sterile. That's the on;y thing I can think of based on what you've posted.


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Invisiblebiologys
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Re: Giving up... for a while [Re: SeniorCoolo]
    #14239069 - 04/04/11 10:21 PM (12 years, 9 months ago)

Quote:

SeniorCoolo said:
You said you wipe your syringe after you flame it. Don't do that. When it's red hot it can't get any more sterile. That's the on;y thing I can think of based on what you've posted.





I agree with this, that is the only flaw i see in your technique, and have you tried several different syringes, from a sponsored vendor? if so that would rule out the syringe part...


how long are you giving to see growth before you're tossing your jars...it can take 2-3 weeks before first signs of growth.


also, are you putting the foil back on after innuculating? tape? dry verm layer?


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OfflineMycelio
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Re: Giving up... for a while [Re: magickpsychonaut]
    #14240334 - 04/05/11 05:50 AM (12 years, 9 months ago)

Quote:

magickpsychonaut said:
I thought that it may have been the syringes, but they come from a reputable vendor and I emailed the vendor who is a highly shroomery recommended vendor who has a very good reputation. I emailed him and  he told me his stuff was good and just said too bad. (I emailed him past the 30 or 90 day guarantee) So I thought it may be something on my part. I'll order from another vendor and see if my results are different.



I think you mentioned Ralphster somewhere. I've seen a lot of complains about moldy spore syringes lately and was sure he would replace them without pretending it was your fault.

Carsten


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Invisiblebiologys
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Re: Giving up... for a while [Re: Mycelio]
    #14246004 - 04/06/11 03:17 AM (12 years, 9 months ago)

i've never had aproblem with ralph not replacing a faulty syringe..


and i've ordered quiet a bit of syringes from him, and only once or twice that i can remember ever getting a dirty one...even then on agar was easily cleaned and able to use.


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OfflinePulseEden
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Re: Giving up... for a while [Re: biologys]
    #14249017 - 04/06/11 06:16 PM (12 years, 9 months ago)

I wouldn't necessarily worry about how clean the bathroom is, it's certainly a great thing that you're cleaning the entire room and it should reduce the chances of foreign spores, but I have done every inoculation I've ever done in a fairly dirty college house kitchen and I have only gotten one small contaminated batch which I attribute to the print I took being contaminated seeing as how the WHOLE batch went bad, and only the batch made with one specific syringe.

Keep in mind, your immediate working area is the most important. The surfaces that are only a few millimeters away from your injection site and the needle of the syringe are really, theoretically, the only things that need to be absolutely clean. The extraneous peripherals are all for the purpose of adding layers of guarantees that those things stay clean. Even the dry verm layer is unnecessary(I still do it).

But there is one possibly important thing you may have overlooked. When you get your syringe, is the needle already attached? If not, make sure when you pull that little cap off of the syringe, and unwrap the needle, that you do so with clean hands. Do it in your glove box, under sterile conditions. I would say that the chances of a contaminant getting in there in the brief moment before you screw the needle on is small, but I wouldn't rule it out.


--------------------
I booped your daughter! A boop boop boop!


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OfflineRedthumb
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Re: Giving up... for a while [Re: PulseEden]
    #14250106 - 04/06/11 09:31 PM (12 years, 9 months ago)

Heres another very important thing to think about. Does your bathroom have an exhaust fan?If it does when you turn on the light and that fan comes on dirty air is being sucked through the bottom and sides of the door. Solution: Use a grow light inside the bathroom and put two clean towels covering the main crack beneath the door.


--------------------


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Offlinemagickpsychonaut
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Re: Giving up... for a while [Re: Redthumb]
    #14257808 - 04/08/11 10:17 AM (12 years, 9 months ago)

@Mycelio, I have done batches with both just flame tipping the needle without wiping with alcohol and also with. Both times I still had contams.

@biologys, all my syringes were from one vendor. I let my jars incubate 8 to 12 weeks. White mycelium does grow, although slow, but it is always contamed with green.

Yes, I am also putting the foil back after innocs, including dry verm layer when making the  jars before sterilization. I do not use tape, instead I have tyvek filters and I have used silicon over the holes in some batches. Always the same contam results.

@Pulse Eden,
so I really don't have to scrub the bathroom like I do? The needles I received from the vendor were not attached. I attached them in a sterile glove box. Do you suggest a trusted vendor that already has the needles attached? I am looking to try at least another vendor to see if my results will be different..

@Redthumb, my bathroom does not have any exhaust fan, and the window stays closed tight. Should I still use a grow light? Also, I will put clean towels covering the crack beneath the door in my next project batch.


THank you all of you for helping me. Hope I didn't ask too many dumb questions


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OfflineMycelio
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Re: Giving up... for a while [Re: magickpsychonaut]
    #14258214 - 04/08/11 12:00 PM (12 years, 9 months ago)

Magicpsychonaut, you spent too much time with a contaminated syringe, always looking for the problem on your side and ramping up your security measures to an insane level. Like PulseEden said, forget about the bathroom and your clothes. Get a clean spore syringe from somewhere else, use your glovebox and it will be fine. After a while, you will even be able to do these inoculations without a glovebox.

Carsten


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InvisibleBoozie
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Re: Giving up... for a while [Re: magickpsychonaut]
    #14258488 - 04/08/11 01:00 PM (12 years, 9 months ago)

Quote:

magickpsychonaut said:
I am also putting the foil back after innocs, including dry verm layer when making the  jars before sterilization. I do not use tape, instead I have tyvek filters and I have used silicon over the holes in some batches. Always the same contam results.



You want to leave the foil off the jars after inoculating. You'll restrict gas exchange otherwise. Also, you don't need tape, tyvek or any form of filtration other than the dry layer of vermiculite [when doing cakes].

I'm not saying these things are the cause of your problems, but keep it as simple as possible and don't divert too far from the proven teks. Keep the tyvek and silicone for grain spawn and just go with a 4-holed lid and dry verm layer for cakes.


What type/size jars are you using?

You said you do see healthy growth initially after inoculating. How long into colonization are things starting to go south? Is it always a green contamination?


Quote:

so I really don't have to scrub the bathroom like I do? The needles I received from the vendor were not attached. I attached them in a sterile glove box. Do you suggest a trusted vendor that already has the needles attached? I am looking to try at least another vendor to see if my results will be different..



I'd keep everything as clean as possible. Clean with soap and water, a vinegar solution, bleach, whatever. Just get it as clean as you can. It can only help. Slacking on sanitation and sterilization is most definitely counterproductive. And ALWAYS use a still-air box (or a flowhood :wink:). Open-air inoculations may work, but why not take extra steps to ensure consistent success?

As for vendors, you'll be hard-pressed to find one that ships syringes with the needles attached. That's not a big deal though. Try Sporeworks. They are the only vendor I use. Well, when concerning spores at least. I've never received a bunk or dirty syringe form them.


--------------------
"After silence, that which comes nearest to expressing the inexpressible is music." - Aldous Huxley


   


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Offlineskiddy
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Re: Giving up... for a while [Re: Boozie]
    #14261778 - 04/09/11 03:21 AM (12 years, 9 months ago)

This makes me paranoid


--------------------
PESH : Pinning
Transeski : colonizing
Orrisa : colonizing
Mex a : colonizing

You're not a mycologist just because you grow mushrooms.


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Invisibleanonjon
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Re: Giving up... for a while [Re: Mycelio]
    #14262010 - 04/09/11 07:10 AM (12 years, 9 months ago)

Quote:

Mycelio said:
Magicpsychonaut, you spent too much time with a contaminated syringe, always looking for the problem on your side and ramping up your security measures to an insane level.




This is what I was thinking.


--------------------
The above post is fictional, hypothetical, or downright nonsensical.


:moon:  :moon:  :moon:  :moon:    :moon:


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OfflinePulseEden
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Re: Giving up... for a while [Re: anonjon]
    #14268704 - 04/10/11 07:41 PM (12 years, 9 months ago)

If the needle is individually wrapped it is sterile. If you attach it in a sterile environment without ever taking the cap off, you don't even need to flame it before inoculation of the first jar. It will be sterile as you take the cap off. Flame it before the second jar of course.


--------------------
I booped your daughter! A boop boop boop!


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OfflineRedthumb
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Re: Giving up... for a while [Re: magickpsychonaut]
    #14272582 - 04/11/11 02:08 PM (12 years, 9 months ago)

Nope, the grow light was just used because the bathroom light in my case is hooked up with the exhaust fan.


--------------------


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Offlinemagickpsychonaut
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Re: Giving up... for a while [Re: Redthumb]
    #14274434 - 04/11/11 07:35 PM (12 years, 9 months ago)

Ok, I have done 5 dummy PF jars. 3 of them I inoculated with a syringe filled with sterilized water in a sterilized glove box. Another jar, I took only the aluminum top off briefly inside the glove box and put it back on. The last jar was a blank that stayed wrapped after PC'ing it.

Here is the step by step:
4. April 2011
1. Pressure cooked 5 PF brf jars for 60 minutes. Cool down overnight.
2. Filled syringe with water, capped needle end, boiled in large pot for 1 hour.
3. Cleaned out entire bathroom, wiped surfaces, walls, counter with bleach.
4. Showered, clean clothes. Waited 2 hours for humidity to dissipate out of bathroom with door open.
5. After 2 hours, Sprayed a few sprays of lysol into bathroom, wiped glove box with bleach and rubbing alcohol, sprayed 2 short sprays of lysol into glove box. Closed box. Closed door, laid towel under door crack.

6. Waited 40 minutes to let settle.
7. Put on sterile gloves. Wiped gloved hands with rubbing alcohol.(Used 70%)
8. Put jars into glove box.
9. Took sterilized syringe out wiped with alcohol soaked pad dry. Flame tipped needle until red, then wiped with alcohol soaked napkin and inoculated each jar. Inbetween each jar the needle was flamed and wiped with rubbing alcohol.

Results: 11. April, 2011 (7 days later) All 3 inoculated jars were contaminated. The one jar that only had the aluminum foil removed was free of any contams. The blank was also free of any contams.



So this clears the air about Ralphsster's syringes being clean. I believe Ralphster's syringes were clean and sterile now.

This rules out pressure cooking as the problem. It must be something else.

Note: In the past I have just flame tipped needle without wiping with alcohol and also did it both ways, with the same results. Also, I have done innoculations both with spraying lysol into the glove box and without.

Any ideas as to what the problem could be? I will be doing dummy jars to rule out one thing after another until I can get to the bottom of things.


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