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Offlineskippydude
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Re: 100% Contam rate. Need advice on filter material. [Re: SmokinErb] * 1
    #17301936 - 11/29/12 08:41 AM (12 years, 1 month ago)

QUOTE
"my lab (PVC frame wrapped in plastic, HEPA intake)."

_____________________________________________________-------------------------

Really surprised none of you vets caught this!

Unless you are using a flow hood or SAB in this setup, it is only semi-contam free until you enter. After entry, no matter how clean you think you are, your lab is contaminated.

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Offlinetwistedty
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Re: 100% Contam rate. Need advice on filter material. [Re: FrankHorrigan]
    #17302256 - 11/29/12 10:03 AM (12 years, 1 month ago)

Quote:

FrankHorrigan said:
Tygard is not tyvek.

I get my SFDs from the mad season.
The 125mm are $1.75 each which is pricey as hell for each jar but I can cut 16 circles from each...comes to 11 cents per lid for a perfect sized filter.





thats what my lid box looks like also, i use exact same tek but with metal, and since i only do agar wedges or g2g i have a single 1/2" hole for teh GE

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InvisibleFrankHorrigan
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Re: 100% Contam rate. Need advice on filter material. [Re: twistedty]
    #17302260 - 11/29/12 10:04 AM (12 years, 1 month ago)

Quote:

twistedty said:

thats what my lid box looks like also, i use exact same tek but with metal, and since i only do agar wedges or g2g i have a single 1/2" hole for teh GE




I made those with inoc ports years ago, I only do agar and G2G as well. But the lids still work nice :thumbup:

I use a 1/4" hole for GE, seems to work great for me.
Any benefit you see from 1/2" hole vs 1/4"?


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

Yes, you can bump my old threads with a question.
Here is how I get things done.
You should take a look. :hehehe:


Frank's tips and tricks. Updated on 3/21/14
AMU- Get an answer here -AMU

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Offlinetwistedty
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Re: 100% Contam rate. Need advice on filter material. [Re: FrankHorrigan]
    #17302287 - 11/29/12 10:09 AM (12 years, 1 month ago)

Quote:

FrankHorrigan said:
Quote:

twistedty said:

thats what my lid box looks like also, i use exact same tek but with metal, and since i only do agar wedges or g2g i have a single 1/2" hole for teh GE




I made those with inoc ports years ago, I only do agar and G2G as well. But the lids still work nice :thumbup:

I use a 1/4" hole for GE, seems to work great for me.
Any benefit you see from 1/2" hole vs 1/4"?




i have seen no difference, ive used both but when i made my lids i followed this video


he uses 3/4" and i thought that was to much.  and at the time i was using 1/4" so i just met in the middle with 1/2"

thats my logic :shrug:

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Offlineskippydude
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Re: 100% Contam rate. Need advice on filter material. [Re: twistedty]
    #17302317 - 11/29/12 10:15 AM (12 years, 1 month ago)

Quote:

twistedty said:
Quote:

FrankHorrigan said:
Tygard is not tyvek.

I get my SFDs from the mad season.
The 125mm are $1.75 each which is pricey as hell for each jar but I can cut 16 circles from each...comes to 11 cents per lid for a perfect sized filter.





thats what my lid box looks like also, i use exact same tek but with metal, and since i only do agar wedges or g2g i have a single 1/2" hole for teh GE




How do you get that red RTV to stick?
Most of the ones I did like that came apart in the PC.
I have to use full size SFD between the lid and ring, with a couple holes in the lid.

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Offlinetwistedty
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Re: 100% Contam rate. Need advice on filter material. [Re: skippydude]
    #17302326 - 11/29/12 10:17 AM (12 years, 1 month ago)

Quote:

skippydude said:
Quote:

twistedty said:
Quote:

FrankHorrigan said:
Tygard is not tyvek.

I get my SFDs from the mad season.
The 125mm are $1.75 each which is pricey as hell for each jar but I can cut 16 circles from each...comes to 11 cents per lid for a perfect sized filter.





thats what my lid box looks like also, i use exact same tek but with metal, and since i only do agar wedges or g2g i have a single 1/2" hole for teh GE




How do you get that red RTV to stick?
Most of the ones I did like that came apart in the PC.
I have to use full size SFD between the lid and ring, with a couple holes in the lid.




grab some sandpaper and rough up the plastic where your rtv or silicone is gonna be

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InvisibleFrankHorrigan
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Re: 100% Contam rate. Need advice on filter material. [Re: skippydude]
    #17302335 - 11/29/12 10:19 AM (12 years, 1 month ago)

Quote:

skippydude said:

How do you get that red RTV to stick?
Most of the ones I did like that came apart in the PC.
I have to use full size SFD between the lid and ring, with a couple holes in the lid.




Sandpaper the plastic around the hole first.
For the inoc ports on those lids you see there, I use a shitload of RTV. Enough to fill the hole and have it bulging on both sides. then i smooth it out and let it cure for a full 24 hours.

If it's not fully cured and you PC it, it will generally fall apart.
Same if you use just a little.
During inoc, you don't want the needle to take the RTV port out with it either, which I've had happen when I used to little RTV.


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

Yes, you can bump my old threads with a question.
Here is how I get things done.
You should take a look. :hehehe:


Frank's tips and tricks. Updated on 3/21/14
AMU- Get an answer here -AMU

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InvisibleFischer
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Re: 100% Contam rate. Need advice on filter material. [Re: RogerRabbit]
    #17306467 - 11/29/12 11:52 PM (12 years, 1 month ago)

Quote:

RogerRabbit said:
Those aren't suitable.  They react with alcohol and allow water/fluids to pass.  We use thicker filters for air.  Just grab synthetic filter disks.  I've never paid over $40 per 100.
RR




Eh?  I might have missed something, but aren't what I posted PTFE (ie synthetic) micron filter discs?

I've never had contamination using them, they shouldn't react with ethanol in mycology applications and the ones I use are .45µm (0.45 micron) pore size, which IIRC is smaller than most/any mold spores and will also not allow bacteria to pass?

They will allow some liquids to pass under pressure (or when wetted with an alcohol, which I didn't realise), but the only liquid that would be relevant here would be water, which will not pass without wetting or pressure and even then will not allow any contaminants that would concern us to pass.

What do you mean by "synthetic filter discs", if not PTFE micron membranes?

Edited by Fischer (11/30/12 12:00 AM)

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Offlineshroom_bee
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Re: 100% Contam rate. Need advice on filter material. [Re: FrankHorrigan]
    #17307154 - 11/30/12 04:41 AM (12 years, 1 month ago)

Quote:

FrankHorrigan said:
During inoc, you don't want the needle to take the RTV port out with it either, which I've had happen when I used to little RTV.




Oh, yeah, f'ing HATE that.  Also, I hate plastic lids.  No matter how rough the sandpaper, or how much I use, the edges of the RTV peel up sooner or later. That shit it not meant to stick together.

I did the same thing as you on the filters, cut them into quarter size circles, and RTVed the hell out of them. Plastic peeled sooner or later, metal, on the other hand, lasted until it rusted away.

Edited by shroom_bee (11/30/12 04:44 AM)

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Offlineskippydude
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Re: 100% Contam rate. Need advice on filter material. [Re: shroom_bee]
    #17307779 - 11/30/12 08:35 AM (12 years, 1 month ago)

You will not know if it is the filter.
Until you do your G2G transfers in a sterile manner.
Doing open air transfers is likely the problem.
             

I have used band aids with 100% success.
When done in the SAB.
IMPO it is not your filter.

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OfflineHioxify
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Re: 100% Contam rate. Need advice on filter material. [Re: c0ri]
    #17308882 - 11/30/12 12:44 PM (12 years, 1 month ago)

I'm in the same predicament.I will most likely  buy them from mycosupply unless I find another source. What are the red dots for, syringe puncture?


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

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OfflineTheCyndicate
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Re: 100% Contam rate. Need advice on filter material. [Re: FrankHorrigan]
    #17308976 - 11/30/12 01:04 PM (12 years, 1 month ago)

Quote:

FrankHorrigan said:
Quote:

skippydude said:

How do you get that red RTV to stick?
Most of the ones I did like that came apart in the PC.
I have to use full size SFD between the lid and ring, with a couple holes in the lid.




Sandpaper the plastic around the hole first.
For the inoc ports on those lids you see there, I use a shitload of RTV. Enough to fill the hole and have it bulging on both sides. then i smooth it out and let it cure for a full 24 hours.

If it's not fully cured and you PC it, it will generally fall apart.
Same if you use just a little.
During inoc, you don't want the needle to take the RTV port out with it either, which I've had happen when I used to little RTV.




In my experience, When making lids with RTV you want to prepare your spawn and get ready to PC your jar's . Right before you load your jars in to your PC, apply the RTV where there is enough on both sides of the lid , put on jar , place band on , Foil and load in to PC.
The intense heat of the PC will automatically cure the RVT , The higher the temp the faster it cures . IMO RTV  sucks it only last like five rounds in the pressure cooker .

Cyn

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InvisibleFrankHorrigan
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Re: 100% Contam rate. Need advice on filter material. [Re: TheCyndicate]
    #17309264 - 11/30/12 01:54 PM (12 years, 1 month ago)

I've been using RTV to seal my filters to my plastic lids for over a year...haven't had to re-seal more than two or three of the hundred or so I've made. I let em cure for at least 24 before PCing.
I do a lot of work too...they've been through countless PC cycles.
I don't even sandpaper the lids first, to be honest. Maybe the RTV fairy just loves me more :fairy:


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

Yes, you can bump my old threads with a question.
Here is how I get things done.
You should take a look. :hehehe:


Frank's tips and tricks. Updated on 3/21/14
AMU- Get an answer here -AMU

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Offlineshroom_bee
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Re: 100% Contam rate. Need advice on filter material. [Re: TheCyndicate]
    #17309273 - 11/30/12 01:54 PM (12 years, 1 month ago)

Quote:

TheCyndicate said:


In my experience, When making lids with RTV you want to prepare your spawn and get ready to PC your jar's . Right before you load your jars in to your PC, apply the RTV where there is enough on both sides of the lid , put on jar , place band on , Foil and load in to PC.
The intense heat of the PC will automatically cure the RVT , The higher the temp the faster it cures . IMO RTV  sucks it only last like five rounds in the pressure cooker .

Cyn




It sucks because you cured it that way. Give it a full 24 hour cure, NO MOISTURE OR PRESSURE, use metal lids, and you will find out the lids rusts from a couple of years of use (20-30 PC runs) before the RTV comes off.

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Offlineskippydude
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Re: 100% Contam rate. Need advice on filter material. [Re: shroom_bee]
    #17309378 - 11/30/12 02:12 PM (12 years, 1 month ago)

Keep the RTV it has cost me one too many LCs.

Full size SFD for me or micro pore tape with heavy duty injection port for grain jugs.

And LC jugs like this

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OfflineTheCyndicate
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Re: 100% Contam rate. Need advice on filter material. [Re: shroom_bee]
    #17309473 - 11/30/12 02:28 PM (12 years, 1 month ago)

Quote:

shroom_bee said:
Quote:

TheCyndicate said:


In my experience, When making lids with RTV you want to prepare your spawn and get ready to PC your jar's . Right before you load your jars in to your PC, apply the RTV where there is enough on both sides of the lid , put on jar , place band on , Foil and load in to PC.
The intense heat of the PC will automatically cure the RVT , The higher the temp the faster it cures . IMO RTV  sucks it only last like five rounds in the pressure cooker .

Cyn




It sucks because you cured it that way. Give it a full 24 hour cure, NO MOISTURE OR PRESSURE, use metal lids, and you will find out the lids rusts from a couple of years of use (20-30 PC runs) before the RTV comes off.




Hmmmm no is sucks because it's not made to be used as an inoculation port. What's this years it takes to rust altered lids. Once you alter and drill a hole in a metal lid and PC them 2-3 times they rust. I noticed this after my first pf-tek.
I have given RTV 24
hours to cure before ..... and it still sucks .
But hey some of my lids still have some RTV ports on them . Done use them anymore ... but they around here somewhere.

Cyn

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Offlineskippydude
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Re: 100% Contam rate. Need advice on filter material. [Re: TheCyndicate]
    #17309504 - 11/30/12 02:34 PM (12 years, 1 month ago)

:handth:

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OfflinePaperTiger
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Re: 100% Contam rate. Need advice on filter material. [Re: skippydude]
    #17309730 - 11/30/12 03:15 PM (12 years, 1 month ago)

Skippy, I like your LC lids. How do the cultures perform? I'm assuming the syringe filter doubles as the gas exchange membrane as well? Any differences in growth, compared to lids with synthetic filter discs and a quarter inch hole? I've never worked with those species, not sure if GE requirements be much different. So when the time comes, you just glove box/flow hood them and quickly install the syringe? How are the contamination rates?

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Offlineskippydude
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Re: 100% Contam rate. Need advice on filter material. [Re: PaperTiger]
    #17309801 - 11/30/12 03:26 PM (12 years, 1 month ago)

Quote:

PaperTiger said:
Skippy, I like your LC lids. How do the cultures perform? I'm assuming the syringe filter doubles as the gas exchange membrane as well? Any differences in growth, compared to lids with synthetic filter discs and a quarter inch hole? I've never worked with those species, not sure if GE requirements be much different. So when the time comes, you just glove box/flow hood them and quickly install the syringe? How are the contamination rates?




For inoculation and drawing cultures I poke a new hole to avoid contamination.

Cultures perform well(have not lost 1 to faulty RTV)

Don't know if it is necessary but have been running 10-20cc air through the whatman weekly for extra oxygen.

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Offlineshroom_bee
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Re: 100% Contam rate. Need advice on filter material. [Re: TheCyndicate]
    #17309986 - 11/30/12 03:58 PM (12 years, 1 month ago)

Quote:

TheCyndicate said:
What's this years it takes to rust altered lids. Once you alter and drill a hole in a metal lid and PC them 2-3 times they rust. I noticed this after my first pf-tek.





Oh, yes, agreed, they will start to rust in a month or 2. But not at the drill points, those are covered with RTV (that seems to stick around for me).  My rims need to be replaced far more often than my lids (6 months or so).  But even the lids usable for years. As long as the myc doesn't mind the rust, neither do I.  Until it goes to the edge and screws up the seal.

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