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OfflineVIgnisFatuusI
Cubist
Registered: 07/07/03
Posts: 50
Last seen: 12 years, 2 months
Question about sterility???
    #1696610 - 07/08/03 04:27 PM (13 years, 5 months ago)

In the PF tek and all other teks I've read, the section on inoculation calls for an extermely sterile envirornment. My question is: Just HOW sterile of a room should you be in to inject spores? The room that we plan to grow in has just been cleaned, but as far as sterilizing goes, how do I know if the air and/or enviornment is sterile enough not to contaminate my substrates? Thanks. Also, when you pressure cook the substrates, obviously, you'd keep the foil wrapped around the jars, but how come in all of the pictures I've seen of pressure cooking demonstrations, I never see any foil?


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Invisiblesoochi
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Posts: 2,420
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Re: Question about sterility??? [Re: VIgnisFatuusI]
    #1696628 - 07/08/03 04:32 PM (13 years, 5 months ago)

I usually pull out the rack in my oven, turn it to 350F for half an hour, then turn it down to 200. place the rack in so its extended as far as it can go, place my jars for innoculation on the rack and innoculate as usual. I also like to spray the immediate area with Oust (air sanitizer) before innoculating. The cleaner you can have your area the better, but don't expect to achieve the sterility of laboratory conditons.


foil is wrapped around the tops of the jars when pc'd then removed after pcing. This is to prevent mositure from getting into the innoculation holes in the lids of the jars.


--------------------
Wee, sleeket, cowran, tim'rous beastie,
O, what panic's in thy breastie!
Thou need na start awa sae hasty,
Wi' bickering brattle!


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OfflinePsilocybin_monkey
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Registered: 06/19/03
Posts: 1,340
Loc: Dragon's Den
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Re: Question about sterility??? [Re: VIgnisFatuusI]
    #1696663 - 07/08/03 04:42 PM (13 years, 5 months ago)

you should build a glove box so you don't have to worry about how sterile everything is in the room.but if your low on money you can do in in front of your oven (oven tek).but if you have to do it in a room for some reason make sure you pick a room with out mats and carpets and you would have to clean the shit out of it and yourself so it's not worth it!So in a few words use flow hood,glove box and oven tek i put them in the order that works the best and cost the most.flow hood really good but cost alot and you may have to build it and to do so you will need alot of tool,glove box is what i use it is cheap to make and easy to build with only needing afew toold that everybody has around the house i have a +air glove box what that dose is forces clean air into the box so no contams can get in."OVEN TEK" all you do is turn you oven on(not sure what exact temp) have the oven door open and place all your sterile tools on the rack that is pulled out for you to work on this cost nothing to do they say the heat forces the air away from the work area and kills contams i never use this tek before i hear people say alot of good things about it and i also hear alot of bad things about it but give it a try if you want it is better to use one of these 3 teks then to clean your room.

NOTE: you could do it in a room that you cleaned BUT the chances of contams are really HIGH (is it worth it?)


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OfflineVIgnisFatuusI
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Registered: 07/07/03
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Last seen: 12 years, 2 months
Re: Question about sterility??? [Re: soochi]
    #1696664 - 07/08/03 04:42 PM (13 years, 5 months ago)

I'm sorry. I think I'm misunderstanding your reply. You said the foil is place while pc'd but removed for pc'ing??? I don't get it. During the process of pc'ing the foil remains on the jar to keep out moisture, right? But after pc'ing, do you remove the foil for colonization? That's what I'm confused about. While the jars remain in the closest for the myceliam to grow, do you keep the foil on? Or does it need O2 to grow? I DID hear, however, myceliam thrives in c02...am I wrong about this?



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OfflineVIgnisFatuusI
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Registered: 07/07/03
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Re: Question about sterility??? [Re: VIgnisFatuusI]
    #1696680 - 07/08/03 04:47 PM (13 years, 5 months ago)

Could someone post a pic of a good glove box construction? That sounds to me like the best, contam-free method. In all of the teks I've read, I haven't seen any with an actual pic of the glove box. I mean, could I simply take a big peice of tuperware, clean it, remove holes for my hands and use THAT as my contam-free environment???? Thanks everyone, again...


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OfflinePsilocybin_monkey
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Re: Question about sterility??? [Re: VIgnisFatuusI]
    #1696803 - 07/08/03 05:31 PM (13 years, 5 months ago)

his is my tek for a good glove box.So far i have had any contams yet (knock on wood)

supplies needed:
1. clear tupperware storage container $5-10
2. 12v computer fan $5-15 (i got a colour fan looks nicer but not needed)
3. gloves $5-15 (the gloves u see are pond gloves)
4. box cutter $1-2
5. 12v power supply $5-20
6. hair dryer or stove $n/a
7. clear plastic $5(table cover or shower curtain)
8. dust mask or coffee filters (hepa filters) $3-5
9. screws for computer fan $1

Making a Glove Box

This is the easiest, effective transfer box I know of. Even if you have to buy everything the cost should be under $35. There are several easy enhancements to this design, but this is a good basic box for beginners and takes about 20 to 30 minutes to throw together.

Construction
1. Cut holes in the side of the box for your arms. Heating the box with a hair dryer or stove makes the cutting much easier.

2. Drill pilot holes to mount the fan to the inside of the box. A hot ice pick works well as a drill.

3. Cut out a hole inside the four screw holes for the fan to draw through. It should be at least 2" in diameter.

4. Mount the fan inside the box, blowing into the box. Seal with duct tape. Put the dust mask on the outside of the box over the draw hole and seal to the box with duct tape.

5. Stick your hand into the gloves. Now stick your arm thru' the arm hole as far into the glove box as you are likely to need to reach. Trim off the excess glove and use duct tape to attach the "glove" to the outside of the box around the arm hole. Repeat for the other arm hole.

6. cut a hole on the top (cover) of the box make sure u make it big because that is what u are going to look through when you are finished i like to leave about 3" around the edge cut using the box cutter it is easier if use heat from hair dryer or hold high over the stove if u do not have a hair dryer then take the clear plastic and put it over the hole cut off extra plastic u don't want it to hang over,then take your duct tape and tape it on.

7. take the 12v power supply with the 12v fan and splice the wires together and use electric tape if u have it (make sure the power cord isn't pluged in while doing this)

8.plug it in and away you go!


(This type of "top of the line" glove doesn't work if you suck air out of the box instead of the positive pressure design, but the best reason to use positive pressure in the glovebox is that leaks in the system are harmless. Filtered air pours out of them instead of unfiltered air being sucked in through them.)

To use: Wipe down (or spray) the inside of the box and the under side of the plexiglas top with Lysol. Spray one glove heavily, then rub your hands (in the gloves) together to clean and sterilize the gloves. Place your materials in the box, lay the top in place, and turn on the fan. Allow it to run for a while (15 minutes is probably overkill). The positive pressure in the box means that filtered air is pouring out of every opening. The dust mask filter can be sprayed with Lysol as an extra precaution.

CAUTION: Using a knife to try to cut the plastic is very dangerous. The polypropylene that these boxes are made of is very tough and will crack without warning if you're using a lot of pressure with a knife. I ruined the first box and could have lost fingers the first time I built one of these. I tried to use a razor knife with a new blade and it did not work. Use a hair dryer and good, heavy duty scissors or tin snips. If you are not experienced with taking power supplies apart and general tinkering, please get someone who is to help with the wiring up of the fan.

i can't get the pictures to load up so use this link to see what it looks like when finished.
http://www.theomc.com/forums/index.cfm?CFID=2569816&CFTOKEN=33040801&fuseaction=read&forum=30&id=381


--------------------
Welcome to my world!



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OfflineSev
Astropath
Registered: 06/06/03
Posts: 1,426
Loc: NY
Last seen: 2 years, 1 month
Re: Question about sterility??? [Re: Psilocybin_monkey]
    #1696846 - 07/08/03 05:43 PM (13 years, 5 months ago)

The oven tek's crap and the glovebox is unnecessary.

Here's what you do:

First, use a tyvek tek. Innoculate through the tyvek.

Second, innoculate in your bathroom. Run the shower on high for about ten minutes, as hot as it goes, so it steams up the room. This will cause most of the airborn dust to clump together and precipitate. Then innoculate.

Third, flame your needle any time that you even think there was a small chance that you might've touched something other than the tyvek and substrate with it. I'd flame between every two jars anyway.


--------------------
"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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OfflineSev
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Registered: 06/06/03
Posts: 1,426
Loc: NY
Last seen: 2 years, 1 month
Re: Question about sterility??? [Re: Psilocybin_monkey]
    #1696854 - 07/08/03 05:46 PM (13 years, 5 months ago)

The oven tek's crap and the glovebox is unnecessary.

Here's what you do:

First, use a tyvek tek. Innoculate through the tyvek.

Second, innoculate in your bathroom. Run the shower on high for about ten minutes, as hot as it goes, so it steams up the room. This will cause most of the airborn dust to clump together and precipitate. Wipe down your working surfaces with isopropyl alcohol. Then, take out your syringe and stuff and innoculate.

Third, flame your needle any time that you even think there was a small chance that you might've touched something other than the tyvek and substrate with it. I'd flame between every two jars anyway.


Sterility is important, but this mostly consists of working fast and touching as little as possible anything that needs to be sterile.


--------------------
"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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Invisiblesoochi
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Registered: 08/13/02
Posts: 2,420
Loc: The Richest County
Re: Question about sterility??? [Re: VIgnisFatuusI]
    #1696863 - 07/08/03 05:49 PM (13 years, 5 months ago)

read my post again. and the oven tek has worked for me so far.


NO! Your CRAP! Muahahahaha!!!!!!!!


--------------------
Wee, sleeket, cowran, tim'rous beastie,
O, what panic's in thy breastie!
Thou need na start awa sae hasty,
Wi' bickering brattle!


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OfflinePsilocybin_monkey
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Registered: 06/19/03
Posts: 1,340
Loc: Dragon's Den
Last seen: 5 years, 1 month
Re: Question about sterility??? [Re: soochi]
    #1696931 - 07/08/03 06:14 PM (13 years, 5 months ago)

no,no,no read my post again hahahaha just messing around

there is still high chance of contams in the bathroom compaired to the glovebox the way i look at it i want to decrease the chance of contams by as much as i can while saving money the oven tek decreases the chance of contams more then it would doing it in a open room and cost nothing!thats shower tek cost nothing too but i think the bathroom is one of the most dirty rooms in the house due to the fact it is bactria heaven but the steam forum water droplets on most/all dust/contams and brings it to the floor but it also dose the same to where you are working and all your tools aswell.the glove box cost under $50 and you spray it with lysol and run the fan for 5-10 mins with the top on/closed to stop any dust/contams from falling into the box and the lysol will kill anything in the glove box and the fan will force clean air into the glove box fo no air born contams and enter the box i do all my work in the kitchen and i spray the work area where i will be sitting my box and stuff down with lysol aswell the reason why i pick the kitchen is that there is no rugs or mats that can hold contams and dust.

but like i said they all cut the chance but the question is how much do you what to cut the chance and how much money you want to spend on it.they way i look at is that if you loss a batch of mushroom due to contams thats can cost you more then what your are trying to save just remmeber a glove box cost under $50 cnd and a batch of mushrooms cost alot more then that to loss to contams!

NOTE:these are just a very rough guess how much it cuts chance of contams and people may do any of these teks and get contam free results it depends on alot such as how clean the tools are?,how clean the house is?,how clean the person is?is there a breeze where the work place is? as you can see there is alot to take into account and these guesses are that everything is as clean as it can get and it may differ for each person.

glove box is like 90+ % contam free - cost $under 50 cnd dollars
oven tek 70+ % contam free - cost nothing
shower tek 40+% contam free - cost nothing

but the differance by 10% or more may lead you to contams that just lost you a $200+ batch of mushrooms so i ask you is it worth it to save a lil bit of money?


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OfflineVIgnisFatuusI
Cubist
Registered: 07/07/03
Posts: 50
Last seen: 12 years, 2 months
Re: Question about sterility??? [Re: Psilocybin_monkey]
    #1697023 - 07/08/03 06:44 PM (13 years, 5 months ago)

Psilocybin Monkey, thank A LOT! That technique seems to be closest to professional sterolization methods. Thanks also for the instructions and pics included. That will become very useful once we begin construction of the glovebox versus simply reading off the tek. Thanks again!


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OfflinePsilocybin_monkey
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Registered: 06/19/03
Posts: 1,340
Loc: Dragon's Den
Last seen: 5 years, 1 month
Re: Question about sterility??? [Re: VIgnisFatuusI]
    #1697044 - 07/08/03 06:55 PM (13 years, 5 months ago)

glad to help


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Offlinefactory81
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Re: Question about sterility??? [Re: Psilocybin_monkey]
    #1697091 - 07/08/03 07:18 PM (13 years, 5 months ago)

Heres my low cost approach towards contam monkey's.


Basic enough as it is, a flowhood. I have thought it over, steralizing a tupperwear box is alot easier then steralizing the whole kitchen. Tupperwear, large kitchen rubbergloves + duct tape = bingo.

Disinfectant wipes with bleach.....hey why not, you have lysol for the air, mys well have wipes for your flowhood, kitchen counters, knobs and handles and what not.

Lysol, spray your air and general surfaces.

Then once youve accomplished all this, use Tyvek or filer discs. I haven't used it yet except for this honey tek jar I just made, but damnit, it sounds convincing. The way things get contaminated is thru the transfer of air and your hands and things. So don't have alot of air movement when your doing your things (sterile glovebox). And with the wipes and lysol you can make yourself add comfort. And then tyvek just is that layer of "little air transfer" for your jars while they iconolate....

All this will cost you like 15-20 bux.


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OfflinePsilocybin_monkey
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Posts: 1,340
Loc: Dragon's Den
Last seen: 5 years, 1 month
Re: Question about sterility??? [Re: factory81]
    #1697211 - 07/08/03 08:18 PM (13 years, 5 months ago)

the air is force out the cracks of the glove box by using a comp fan that has a filter over it such as hepa,coffee,dust mask of the thus no dirty air what so ever gets in the box that is the whole point!!! and to make sure there is no dirty air already in the glove box it is sprayed with lysol killing all contams

a flow hood for $20 hahah!!! that tek is not a flow hood what so ever flow hoods cost $hundreds of dollars and they use a power fan and a hepa filter here's some links

http://www.thelittleguy.com/flow_hood.html

http://www.theomc.com/index.cfm?fuseAction=teks.lsosFlowhood


--------------------
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OfflineVIgnisFatuusI
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Registered: 07/07/03
Posts: 50
Last seen: 12 years, 2 months
Re: Question about sterility??? [Re: Psilocybin_monkey]
    #1704070 - 07/10/03 10:58 PM (13 years, 4 months ago)

What about a glove box transfer in a steamed bathroom!?!?! Best of both worlds?


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