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Offlinekyleda1kid
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pf tek success rate?
    #7856862 - 01/09/08 11:10 PM (16 years, 2 months ago)

Using pf tek method with 1/2 pint brf jars pressure cooked 45-60 min. cooled overnight. Of course follow 2:1:1 ratio (verm:brf:water) then inoculated with spores from ralphsters using a glovebox. I am only getting a success rate of about 60-70%; 6-7 out of 10 jars are good. Others get some sort of contaminate. I spray Lysol 20-30 min before. I use hand sanitizer, sterilize needle of syringe every 2-3 jars with lighter, and still something is wrong. Am I missing something, I know they all can't always work but 60% success rate? Is that normal? By the way pressure cooker is only a 6 qt. holds exactly 8 jars. Don't know if that matters.

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InvisibleKillerPicklez
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Re: pf tek success rate? [Re: kyleda1kid]
    #7856997 - 01/09/08 11:28 PM (16 years, 2 months ago)

Quote:

kyleda1kid said:
Using pf tek method with 1/2 pint brf jars pressure cooked 45-60 min. cooled overnight. Of course follow 2:1:1 ratio (verm:brf:water) then inoculated with spores from ralphsters using a glovebox. I am only getting a success rate of about 60-70%; 6-7 out of 10 jars are good. Others get some sort of contaminate. I spray Lysol 20-30 min before. I use hand sanitizer, sterilize needle of syringe every 2-3 jars with lighter, and still something is wrong. Am I missing something, I know they all can't always work but 60% success rate? Is that normal? By the way pressure cooker is only a 6 qt. holds exactly 8 jars. Don't know if that matters.




how many times have you got this success rate? I know my first 1-2 times, my success rate was at like 40%, then 60%, then finally I figured my shit out and got consistently over 75% success.

my errors seemed to be at the inoculation point. I learned that it is best to just re-sterilize the needle after each jar. takes a couple minutes extra, but seemed to be worth it.

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OfflineTHEBats
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Re: pf tek success rate? [Re: KillerPicklez]
    #7857014 - 01/09/08 11:31 PM (16 years, 2 months ago)

So far I have had 100% success and I used only steam sterilization. Out of 6 grows that can't be too bad.

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Offlinekyleda1kid
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Re: pf tek success rate? [Re: KillerPicklez]
    #7857018 - 01/09/08 11:31 PM (16 years, 2 months ago)

Only 3-4 times at most. I'll try re-sterilize after each jar. Thanks, any other suggestions?

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InvisibleKillerPicklez
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Re: pf tek success rate? [Re: kyleda1kid]
    #7857025 - 01/09/08 11:32 PM (16 years, 2 months ago)

Quote:

kyleda1kid said:
Only 3-4 times at most. I'll try re-sterilize after each jar. Thanks, any other suggestions?




Hm. Are you sterilizing the jars after each grow? And where do you store the jars during colonization?

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Offlinekyleda1kid
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Re: pf tek success rate? [Re: KillerPicklez]
    #7857055 - 01/09/08 11:36 PM (16 years, 2 months ago)

Of course I re-sterilize jars, wash with bleach water then wash with soap and water. I keep jars in a rubbermaid container with lids slighltly open. They are kept in a dark place 72-80 degrees. There is also holes in the rubbermaid container to help with gas exchange.

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Offlinekyleda1kid
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Re: pf tek success rate? [Re: kyleda1kid]
    #7857066 - 01/09/08 11:38 PM (16 years, 2 months ago)

i meant re-sterilize the needle not jar other post

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Re: pf tek success rate? [Re: kyleda1kid]
    #7857159 - 01/09/08 11:52 PM (16 years, 2 months ago)

sounds like you do everything perfectly, i can see no holes in your procedure whatsoever.

dont know what advice to give you. they are getting contaminated during colonization right? not in the fruiting chamber.

are you using distilled water in your substrate preparation?

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Offlinec0_hush
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Re: pf tek success rate? [Re: KillerPicklez]
    #7857538 - 01/10/08 01:41 AM (16 years, 2 months ago)

Couple of suggestions and a question.. In my experience, having the jars wait longer than you should in the PC increases the chance of contams IMO... I always inoculate 5 hours after I'm done PC'ing. Also, how thick is your dry verm layer? Make it at least 1/2 inch thick cuz it's your only defense against contams after inoculation. How much water are you using the the PC? Too much water will get in your jars and mess up the dry verm layer and/or the water ratio and could lead to contams.. Flame the needle after each jar just to be on the safe side and PC for AT LEAST 60 mins, if you're getting contams, do it for 75, to play it safe and rule out stuff.


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Offlineatomicblue
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Re: pf tek success rate? [Re: c0_hush]
    #7857577 - 01/10/08 01:54 AM (16 years, 2 months ago)

100% for me too. First did nine jars now another 10. All are fine...

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Invisiblefastfred
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Re: pf tek success rate? [Re: atomicblue]
    #7857686 - 01/10/08 03:34 AM (16 years, 2 months ago)

Detail your contaminations. Locate where they are coming from.

After a couple runs you should be a lot smoother and quicker in your innoc technique. You should be getting +90%. 98-99% is not hard to obtain in most cases.


-FF

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OfflineNibin
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Re: pf tek success rate? [Re: fastfred]
    #7857693 - 01/10/08 03:53 AM (16 years, 2 months ago)

I have made over 100 pf cakes all steamed, not pc'd and not one contaminated. Something is wrong.

Was the syringe made by you from a print by ralphsters od did you buy the syringe directly?

I suggest you cover your hair and mouth when innoculating, or even better, use a still air box or glovebox.

Also, check if your dry verm layer is getting wet during the sterilization as that would render it useless.


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Re: pf tek success rate? [Re: Nibin]
    #7857738 - 01/10/08 04:51 AM (16 years, 2 months ago)

> check if your dry verm layer is getting wet during the sterilization as that would render it useless.

Something has to be useful in the first place before it can be rendered useless.

It's not really possible to heat a moist substrate well above boiling and not have a dry, water-absorbent layer directly above it become moist.

We need more details in order to locate the source of your failure. Where did the contamination occur? At innoc sites?


-FF

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Re: pf tek success rate? [Re: fastfred]
    #7857778 - 01/10/08 05:42 AM (16 years, 2 months ago)

At first i had a poor success rate, 3/4 jars contaminated. But since moving to a glovebox. 100%.

Dry verm layer, thats a good point. It must be dry.

I noticed you say sterilised for 45-60 minutes.

I will maybe suggest pressure cooking a little longer. 60 minutes min. I always give mine an extra 15 minutes just to be sure. The pressure cooker shouldnt be a problem, i use a small 1 that holds 4 1/2 pints (8 if stacked)

You should flame the needle between each jar to prevent cross contamination. So flame before you inoc each jar.

If they are properly sterilised then infection can be happening during inoculation.


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InvisibleDontPlay
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Re: pf tek success rate? [Re: veda_sticks]
    #7857785 - 01/10/08 05:47 AM (16 years, 2 months ago)

Make sure you are cooking them for at least 60 min not 45...Second are you sterilizing the needle between each jar, you should even in a glovebox small things will make all the difference. My first dozen jars I made had only 1 contam...And I know it was my fault because I was messing with the foil, and I had no dry vermiculite barrier.


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Offlinec0_hush
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Re: pf tek success rate? [Re: veda_sticks]
    #7857787 - 01/10/08 05:48 AM (16 years, 2 months ago)

I pretty much just make sure to do a full 60 min PC as stated above and flame the needle after inoculating each jar, all in open air. I ditched my glove box completely and don't miss it.


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Re: pf tek success rate? [Re: DontPlay]
    #7857917 - 01/10/08 07:16 AM (16 years, 2 months ago)

Quote:

DontPlay said:
Make sure you are cooking them for at least 60 min not 45...




20 minutes is the standard sterilization time for PF jars, and that's at 12 psi. Sterilizing for 30 min at 15 psi provides a more than adequate safety margin. Sterilizing for 45 min is acceptable, but any longer is badly overcooking your substrate. You are simply breaking down nutrients, caramelizing sugars, and wasting time/energy.

The above applies to elevations below 2000 feet. For higher elevations calculate your temp at 15 psi, then double your sterilization time for every 10C difference.


-FF

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Re: pf tek success rate? [Re: veda_sticks]
    #7858128 - 01/10/08 08:28 AM (16 years, 2 months ago)

Quote:

veda_sticks said:
At first i had a poor success rate, 3/4 jars contaminated. But since moving to a glovebox. 100%.

Dry verm layer, thats a good point. It must be dry.

I noticed you say sterilised for 45-60 minutes.

I will maybe suggest pressure cooking a little longer. 60 minutes min. I always give mine an extra 15 minutes just to be sure. The pressure cooker shouldnt be a problem, i use a small 1 that holds 4 1/2 pints (8 if stacked)

You should flame the needle between each jar to prevent cross contamination. So flame before you inoc each jar.

If they are properly sterilised then infection can be happening during inoculation.




Same here everything I grew was green until I started inoculation in still air,now I hardly get contamination at all

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Offlinegaladar


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Re: pf tek success rate? [Re: Blutjager]
    #7858198 - 01/10/08 08:45 AM (16 years, 2 months ago)

tips to stay good for the pf tek.
-wipe lip of your jars. a good 1/2-3/4 inch
-have some micropore tape over holes, just so you create the smallest possible hole with the needle.
-flame between every jar and stick it straight in the jar while red hot.
-use a gloveox
-make a 10:1 bleach and water spray and youll be good. spray this shit everywhere. the air, the surfaces, alot inside your glovebox lol. i wear something over my mouth and nose so i dont inhale like a dumbass and burn my throat. works better than any lysol and has no smells. well a little. i like 9:1 better.

and dont know if this helps, but i micropore tape again after inoculation. and then whel my cakes are 50% colonized( a bit stronger) i take off all tape.

100% rate here about 30 pf jars in
grains is another success story =]

Edited by galadar (01/10/08 08:57 AM)

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OfflineGrylls
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Re: pf tek success rate? [Re: galadar]
    #7858232 - 01/10/08 08:54 AM (16 years, 2 months ago)

I don't have the luxury of a glovebox, so I wipe everything with alcohol: my hands, jars, syringe, alcohol bottle, counter top and the lighter. I use latex gloves and wipe them down too before each jar. I flame the needle for each jar and cover the hole with a cosmetic cotton pad moistened with alcohol right before insertion. I replace pad and get a band aid ready to stick over the hole. I then put the jars into a semi-sterile environment for incubation.

So far, my first 5 jars have been 100% free of contaminants. I just knocked up 5 more. Let's see how they go.


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