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Amazon Shop for: ½ Pint Jars, Agar, Autoclave, Impulse Sealer, Mycobags, Rye Grain, Wild Bird Seed

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OfflineTweexican
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some help refining my proceedure
    #3432510 - 12/01/04 01:53 PM (12 years, 4 days ago)

I have successfully colonized both wbs flour cakes, popcorn jars, millet jars, and rye jars. My problem, however, comes mostly from contam within my jars. Mycellium will grow, but then the jars usually contam. I really don't know what's causing it.

I use tyvek, cut in 4"x4" squares on top of the canning lid, held in place by the metal ring. (I'm using quart jars of course.) My biggest concern is that I'm using the tyvek wrong, (either wrong side up, or in place wrong). My proceedures are pretty sterile, I work in front of a flowhood.

Could anyone help? :sad:


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Offlinespliffmasta
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Re: some help refining my proceedure [Re: Tweexican]
    #3432526 - 12/01/04 01:57 PM (12 years, 4 days ago)

Hrmm, I don't know whether it is a greater risk of contamination, but I've found that not using the discs in the lids, with just tyvek and the lid causes the substrate to dry out significantly faster/easier. I'd suggest 5 x 1/4" holes in the disc and do both tyvek and the disc.

Are you handling your jars too much? I find that I get more contams the less patient and more curious I get.


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InvisibleTODAY
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Re: some help refining my proceedure [Re: Tweexican]
    #3432530 - 12/01/04 01:59 PM (12 years, 4 days ago)

are you pc'ing your grains? i pc my wbs and only use alluminum foil to cover the lids during pc'ing and another layer to cover the innoc point and get barely any contams. check the sterility procedures you use while printing and syringe making.


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OfflineTweexican
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Re: some help refining my proceedure [Re: spliffmasta]
    #3432535 - 12/01/04 01:59 PM (12 years, 4 days ago)

i use the metal disk with 4 1/4" holes (not specific) then the tyvek, then the metal lid.

do you cut your tyvek into disks or squares?

I use squares and just tighten them on with the metal ring of the jar.

(grain is prepared by 24hr soak, rinse, 30min simmer, pc at 15psi for 1hr)


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Edited by Tweexican (12/01/04 02:01 PM)


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Invisibleagar
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Re: some help refining my proceedure [Re: Tweexican]
    #3432541 - 12/01/04 02:00 PM (12 years, 4 days ago)

TYVEK goes on grain or WBS jars like this.







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Offlinespliffmasta
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Re: some help refining my proceedure [Re: agar]
    #3432546 - 12/01/04 02:02 PM (12 years, 4 days ago)

agar, dropped you a PM, hook us up with a nice tek post on those things.  :wink: :heart: :thumbup:


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OfflineTweexican
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Re: some help refining my proceedure [Re: agar]
    #3432548 - 12/01/04 02:03 PM (12 years, 4 days ago)

agar, how do you cover your inoculation points?

I am pretty sure the tyvek goes on TOP of the lid... anyone else?


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Invisibletripndicular
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Re: some help refining my proceedure [Re: Tweexican]
    #3432556 - 12/01/04 02:04 PM (12 years, 4 days ago)

Have you considered altering the amount of water you use in each mix ?
Try cutting back a tad on the water , too much moisture can be a problem .
Also are you using tap water or distilled ?
Make some "minor" adjustments in your procedre and see if it helps .


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OfflineTweexican
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Re: some help refining my proceedure [Re: tripndicular]
    #3432564 - 12/01/04 02:06 PM (12 years, 4 days ago)

most of my contams end up starting at the tops of my jars... The mycelium will germinate, colonize for a few days, then im stuck with a war in the jar. I seriously have like 10 jars that I'm holding onto with a little contam spot at the top with the rest 70-80 percent colonized. thats why I am wondering if maybe it is because of complications with the lid...


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Edited by Tweexican (12/01/04 02:08 PM)


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Invisibleagar
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Re: some help refining my proceedure [Re: Tweexican]
    #3432575 - 12/01/04 02:09 PM (12 years, 4 days ago)

On metal lids with Tyvek, you can drill 2 holes & innoc through one, then tape over.

Or....drill 3 holes. Two ....1/4 inch towards the edges & across from each other.

Center, drill smaller (1/8th or 3/16th) hole for injection site.

Inject in center hole, then just alc swab tape & tape over.

Simple as that.


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OfflineTweexican
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Re: some help refining my proceedure [Re: agar]
    #3432599 - 12/01/04 02:16 PM (12 years, 4 days ago)

shit, I'm getting sick of jars. They're just a hassle. Has anyone here switched from jar to mycobag?


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Invisibleagar
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Re: some help refining my proceedure [Re: Tweexican]
    #3432919 - 12/01/04 03:40 PM (12 years, 4 days ago)

Not Mycobags, but.....autoclave bags, w/filter patch & impulse sealer. They are GREAT...for lots & lots of spawn. Large All American PC the bigger the better is great for doing them.


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InvisibleMagashM
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Re: some help refining my proceedure [Re: Tweexican]
    #3432934 - 12/01/04 03:43 PM (12 years, 4 days ago)

Are you making your own syringes. I do my jars exactly like you do.


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Invisibleagar
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Re: some help refining my proceedure [Re: Magash]
    #3432992 - 12/01/04 03:59 PM (12 years, 4 days ago)

Quote:

Magash said:
Are you making your own syringes. I do my jars exactly like you do.




Are you asking ME?

If so, YES..... I make my own prints, syringes, & liquid culture.


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OfflineIGnosticAbhorI
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Re: some help refining my proceedure [Re: agar]
    #3433026 - 12/01/04 04:07 PM (12 years, 4 days ago)

Not sure if it'll help put I put coffee filters over my lids. I have polyfil and i put two coffee filters of that and secure it down with a rubber band. Someone suggested it for really dirty places, like attics and basements, my procedures are pretty sterile and now that i have polyfil and coffee filters i've yet to get a contam. and i have a Big contam. problem awhile ago. Well gl, hope ya find something
Later


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Offlinerecalcitrant
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Re: some help refining my proceedure [Re: agar]
    #3433057 - 12/01/04 04:15 PM (12 years, 4 days ago)

Quote:

agar said:
Are you asking ME?





You can (sometimes) tell where the post was directed by checking the [Re:USERNAME] that follows the post title.

Using Magash's post as an example we see that the post titles reads
Quote:

Re: some help refining my proceedure [Re: Tweexican]
#3432934 - Wed Dec 01 2004 12:43 PM




So, it's likely that Magash clicked either the "Quick Reply" or "Reply" link in the post title of Tweexican.

Checking to whom the reply was addressed doesn't always work because a hell of a lot of people are oblivious to this feature.


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Offlinemidway
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Re: some help refining my proceedure [Re: IGnosticAbhorI]
    #3433062 - 12/01/04 04:17 PM (12 years, 4 days ago)

i get 95% to 100% colonization in two weeks using pints of millet with the metal ring and covered with foil...ie pftek style. Ive never used tyvek, but im pretty happy with my colonization speeds without it, and frankly im too contam paranoid to try it. Maybe quit using it and see if that helps? Maybe its overrated...


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Amazon Shop for: ½ Pint Jars, Agar, Autoclave, Impulse Sealer, Mycobags, Rye Grain, Wild Bird Seed

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