|More talk...||10||02/28 08:13am|
|Gloveboxes & Sterile Technique||-|
|Incubation after Inoculating||-|
|Step By Step Pictorial w/text||3||11/16 08:32pm|
|Overkill in flame-sterilizing??||5||01/15 08:02pm||Hatcher|
|By Nan (Nanook) on Thursday, October 18, 2001 - 05:14 pm:|
I prefer to innoculate jars and do Glove Box work in a "Clean Room", and I like to use the bathroom because it needs to be cleaned periodically in any case and it is small: less to clean.
I use clorox and soap then rinse with 10% clorox on a clean cotton rag. Floors, sink, tub, toilet paying special attention to the door knobs, light & fan switches and the faucets. Remember bathrooms should be well cleaned all the time anyway, just do an extra good job and use clorox solution to wipe.
After everything is wiped and still damp I move in my cooled pressure cooker and set it in the tub, glovebox on the sink if I am going to be using it, then I make sure the vent is closed and the fan is off when leaving. And I let the room sit and dry for 20-30 minutes with the door closed, it gives the clorox a chance to work.
I have found that taking a shower and wearing clean shorts before innoculating cuts down on the incidence of trich, or green mold in shroom cultures.
Then I go in and sit down, light a small alcohol burner to flame needles, and go to work. My exact handling of the jars is covered in Foil Covers at the bottom.
|By Municipal Jones (Phishgrower) on Wednesday, September 19, 2001 - 11:00 pm:|
Hey, 2 more questions here. First, the only contamination I'm worried about for my second attempt is during innoculation. This is what I'm planning on doing: Heating my stove at the lowest setting and then placing the jars I plan to innoculate on a level in the oven. I've read that doing it over heat like this will prevent contams. Is this a good method or are there much better methods? Also, with all hopes my second batch will actually produce results. Assuming I'm able to grow what advice does anybody have on taking spore prints? What's the easiest full proof method of doing this so I don't have to keep shelling out money for the syringes? Also, can you reuse a syringe if you have a viable solution? Thanks.
|By saluras (Saluras) on Wednesday, September 19, 2001 - 11:24 pm:|
here we go:
1. i've never had a problem just innoculating the jars in an open room. i've posted about it, my place is a crappin mess, i've done it with the air running, with the windows open, cigarette smoke in the air, never any contams with colonization (3rd or 4th flush yeah, but what do you expect).
until recently i've been making prints in the pint jars. i sterilize a jar for 30 minutes. i then cut (or pull) the cap off the shroom and put it into the jar. instead of the lid i put a piece of thick paper towel over the jar and twist the lid on. after about 24-48hrs there is a nice dark purple print. I usually then fill the jar with sterile water and use for syringes.
i haven't paid for spores in a long time, i only recently started trading. if you follow this you will have good ususable syringes every time.
its kinda strange, because i hear about how many problems people have with this, but i swear i just don't have problems. also those syringes with 24hr old spores colonize fast.
hope this helps
|By monkeyod (Monkeyod) on Saturday, September 22, 2001 - 03:28 pm:|
The Oven Tek is a good way to go, it creates an updraft of warm air that will keep anything from falling into your jars. Just flame your needle before injecting and flame it again if touch anything (besides normal). Just keep it as clean as possable and all should be fine.
|By trippysmurf on Monday, August 13, 2001 - 12:37 pm:|
thanks nan, good advice, my jars are currently in my glove box, and ill be covering them with foil again tommorow, can i just cover the holes with foil and tape to save on foil? or would it be better to cover the whole top? sory nan just want it perfect. first signs of mycellia are pretty exciting time huh? well for newbies like me i guess
|By Nanook of the North (Nanook) on Monday, August 13, 2001 - 09:03 pm:|
Trippy: If you wanted "perfect" or as close to perfect as possible... Then when inoculating jars work in a glove box, remove the foil cap, place it on a sanitized plate, peel up the tape, shoot the holes, press the tape back down, and replace the foil cover (which was never turned over). Place the jars in a clean, draft free, location and let them sit undisturbed with temps in the 80's until they are colonized... That is pretty close to perfect.
I do not use tape, and once in awhile I will find some cobweb in the dry verm layer if it's dusty. I don't use a glovebox. I cover my jars with a foil cap, and only remove the foil long enough to shoot the jar, then it is replaced. I inoculate in a still sanitized bathroom. Store the jar in a clean warm cabinet with the doors closed.
Leaving the foil caps on is a good habit to get into BTW. It keeps the rings clean too, which is something you want, especially with Dextrose jars.
|By CaptainMaxMushroom on Tuesday, August 14, 2001 - 01:30 am:|
You know guys, its really a bunch of overkill. You don't need holes, tape, foil, none of it!
Common sense would have you working in a clean area to begin with right? Just take the lid off, quickly innoculate then put lid back on. It's that simple.
I don't know why people think that by default, the air is contaminated. It isn't! Not unless you live in a morgue or leper colony!
I never use foil, holes or tape and don't have any problems at all. If the syringe is clean, you shouldn't have any problems either.
Visit the SporeLab @:
It's a trip you'll never forget!
|By trippysmurf on Tuesday, August 14, 2001 - 03:41 am:|
actually, the only sterile procedure i did was making the syringe in the glove box, then i thought about the possible points of contamination, and i read somewhere on the old board that "you could hock a loogey in the jars before sterilization and it wouldnt matter" not that i would, but i figured that if i was gonna sterilize, bacteria wouldn't matter while mixing up my substrate, the only point of contamination would be innoculation, from airborne contams, but shouldn't the verm barrier take care of that? so basically what the foil does is to keep bacteria from entering the holes right? wouldn't the micropore tape be enough? although i know you guys are right as what we are going for here is the optimum setup, i was just wondering what corners i could cut pretty arrogant for a newbie huh? well everythings peachy, and when things go sour, i know ill be running back for help! hahaha...
(soon to be papasmurf!)
|By Nanook of the North (Nanook) on Tuesday, August 14, 2001 - 04:09 am:|
Start real real clean... Then get sloppy...Until your first batch of contams show up, then get clean again Once you are more familiar with it, you learn where you can cut corners and where you can't.
But if you start out using good sterile technique your contams will be few and far between.
|By Poopypants (Poopypants) on Tuesday, August 14, 2001 - 08:39 am:|
Making Jars and such, I don't bother being too sterile. I've always gone by the theory that if it gets Contammed, it was a weak. With the amazons I have now - if I get contams I just chop it out... it overgrows the spot. The less you baby , the less chance you have of developing a wimpy strain that dies in the dozens every time you sneeze.
On the flip side - when I do Prints - Agar - Syringes etc. I'm extrememly Sterile, I wash me and evrything within a mile with alcohol. If i'm going to get contammed I know exactly where it came from.
|By onediadem (Onediadem) on Saturday, November 03, 2001 - 02:08 am:|
I ordered 2 syringes from PF. When they arrive, what are they packaged in, and do I need to not open pkg until ready to inoculate? I have never ordered syringes before. Should I take them out of pkg, and put them in my glove box? Can you spray the outside of the covered syringes w/lysol? Or post a link for this? I searched and couldnt find what I was looking for.
Nan, I believe I will be using the karo syrup tek, looks good, and I like the idea of getting more for your buck.....thx
keep it loose, pam
|By Lichen (Lichen) on Saturday, November 03, 2001 - 02:19 am:|
the syringes come in a cardboard box 2"x2"x10" and they are wrapped in newsprint. They are sterile inside, and clean enough outside. Flame sterilize the needles and you'll be fine
|By Nan (Nanook) on Saturday, November 03, 2001 - 02:32 am:|
Open the box and check them out, then close it up and store it in the fridge until you are ready to use them. You can spray the outside of the syringe with Lysol or alcohol, make sure they are completely dry before use...
If I am shooting liquid spores into dextrose I sterilize my jars and utensils in the PC: some extra jar lids wrapped in foil (empty syringes if I need them), a plate, some extra foil to make a little sterile tray to hold syringes and instruments.
Clean the box. Open the PC and load your cooled jars, extra lids, whatever into the glovebox. Remove the tape guard from the needle cover on the syringe, wipe it down with alcohol and place it in the glovebox. Mist everything lightly. Let dry. Begin glove work.
You'll do fine, just anxious.