|Archives -> Spore Printing||-|
|Print Question||15||12/31 03:11am||Kevin Smith|
|Gill Development and Spore Prints....help||12||01/06 06:31am||Kevin Smith|
|My first print success||12||03/01 08:08pm||Scotsman|
|By Lichen (Lichen) on Thursday, November 01, 2001 - 03:45 pm: The Nook|
I am determined to take a spore print of my B+ before it's too late; and I happen to have a cake that has three fine specimens on it. The question is, do you all think I can use a shroom whose cap has turned upside down? I am afraid that being upside down, it is possible some kind of resident contaminant has perhaps settled on the gills. So should I use one that it right side up, or can I use the upside down one? (The only reason I want to use it is because it is nearest to maturity)
|By Nan (Nanook) on Thursday, November 01, 2001 - 04:05 pm: The Nook|
Use a normal shaped cap.
|By relic (Relic) on Thursday, November 01, 2001 - 04:48 pm: The Nook|
yeah, i wouldn't use one that is turned upside down. i think one of the reasons prints are so clean is because of the umbrella effect of the cap.
|By Lichen (Lichen) on Thursday, November 01, 2001 - 06:23 pm: The Nook|
This is what I anticipated you'd say...the umbrella effect will keep contams from settling on the gills. And Nan's betadine trick might come in handy, too. I wish I could say I've done this successfully before, but I'm still trying
|By Fishy1 (Fishy1) on Friday, November 02, 2001 - 01:45 am: The Nook|
To make it easier, i stab the cap with a syringe, cut the stem, then the cap is held, inpaled on the needle. Use the "V" of the open scissors to slide the cap off the needle and onto the paper (or foil...). Pop a sterile bowl over them-done.
BTW- I just took some GTs and TCs.......
|By Lichen (Lichen) on Friday, November 02, 2001 - 01:59 am: The Nook|
I used a straight pin a la pftek...but I know for certain that shooting dozens of jars with contaminated spores will be simply heartbreaking. Oh, well, I MUST have success at this or I'm not really a mycologist (even an ameteur, hobbyist mycologist needs to be able to collect viable spores)
|By Nan (Nanook) on Friday, November 02, 2001 - 02:10 am: The Nook|
Lichen, before you shoot dozens of jars... Shoot one jar of Karo Tek and give it 5 days to germinate. You will know if the spores are clean _before_ you go through the effort of making up and shooting jars...
This implies that Karo Tek works! Yay! It _works_ guys, all my liquid cultures will be ready to shoot tomorrow or the day after at the rate they are progressing.
I will write up the Tek tonight and get it posted.
|By Lichen (Lichen) on Friday, November 02, 2001 - 05:07 am: The Nook|
I saw the tek, Nan. It looks very nice. Alas, I shot several jars with the microwave honey tek today. We shall see. I'll try the karo with my B+ spore print in a day or two. We'll see if I managed it soon enough.
Then if I can save the albinos that I have, too. Did you find your great white whale, Nan?
|By Nan (Nanook) on Friday, November 02, 2001 - 05:24 am: The Nook|
Not yet. Ahab's quest went through several straw beds... And lots of aborts... Without finding the Great White... A few pretty ones, but no giant... Yet! I needed inoculum... Planned to shoot another Dextrose jar with Albino, and got sidetracked with the Karo tek... So now I have 400 cc's of Hawaiian clone that will need to be used up... Not that I am crying in my liquid culture mind you, this clone fruits giant 40-45 gram shrooms on straw... Two dried shrooms weigh over a 1/4 ounce... And I get 15-20 of them on a 2 sq foot bed. I'll find the White Whale though, this search will just take time.
|By Hatcher (Hatcher) on Friday, November 02, 2001 - 09:21 am: The Nook|
Good god Nan, two dried shrooms weighing over a qtr Z? And still looking for bigger fish? Ever think Ahab might have been a madman?...:-)
Lichen, it's no sweat, I got it right the first time, all the way to my own syringes. But first the print. I used a half-pint jelly jar, the inoculum made in same. First question, do ya know how to use your oven, aka I think, as a "flow hood"? Open the door no further than to pull the middle rack out enough to create a work area, and turn it on, LOW. A slow gentle heat is all that is needed. Having killed all fans, central air, whatever, freshen the air w/Lysol-type areosol, and wipe down everything w/alcohol. The heavy-duty aluminum foil for the oven rack, kitchen counter, scissors, all of it. I straightened out two large safety pins to move the cap, one to stick it, one to push it off. When your choice is dropping it's spores, (wait to see that), drop a sandwich baggie over it, pluck it, and take it to the oven. Just don't breathe on the work, wash those hands, everything should be fine hoss. (I'm assuming you can fill in a few blanks!) Screw it down tight, 24hrs. is gonna be plenty, get your kitchen right again, remove cap, leave the lid screwed on kinda loose for about a week, then tighten 'er down until needed. I've got four outta five races sittin' in jars just waitin'. Only made one jar of inoculum so far, all I've needed.. A little over a quarter pint goes a long way. But even from the same jar of juice, I've still been making the syringes, then testing each for contams by inoculating each to a corresponding jar. So far tho, so good! And pretty simple.
Hope this helps you out Lichen!
(good enough for the big A's Nan?)
|By Nan (Nanook) on Friday, November 02, 2001 - 09:30 am: The Nook|
Yep, when this thread goes stale I'll add to the spore printing folder in the Archives.
Thanx for the post!
|By Hatcher (Hatcher) on Friday, November 02, 2001 - 09:37 am: The Nook|
You're welcome Nan, I thank you dude. Peace!
|By SYDYSTYK (Addict) on Friday, November 02, 2001 - 11:46 am: The Nook|
the jars in the oven work for me too, only messed up one so far, be sure to make a honey/dex/karo jar, i lost a ton of jars due to 1 bad print infected with Cobweb
See: Contams in Liquid Culture
|By Lichen (Lichen) on Friday, November 02, 2001 - 02:35 pm: The Nook|
I'm not getting excited yet, but I am gaining in confidence. Like I said, we'll see soon
Thanks, Hatcher. I never have used the oven, but I understand the concept.
I will make a glovebox, a glovebox is what I need
[Indeed... See: Lichen's Glovebox]
|By Hatcher (Hatcher) on Friday, November 02, 2001 - 05:27 pm: The Nook|
That's the point of using the oven, eliminates Glove Box. The rising warm air keeps comtams off. I wonder how these guys get the work in and out of glove boxes w/o contaminating the airspace inside--gotta have a Clean Room too? Tellin' ya dude, use the oven and preserve your races...
|By Nan (Nanook) on Friday, November 02, 2001 - 05:40 pm: The Nook|
I will tell you how I do it in mine...
I put all my stuff in the box and spray everything down with Lysol. Then I turn the fans on and let it sit for 20-30 minutes until the Lysol has dried and the fumes have dissipated. Then I pop on some gloves, wash them with a few mils of alcohol, and go to work in the sterile box. I work in a bathroom, which is the cleanest room in this house.
A cheap effective box is worth the time building. I hate slaving over a hot stove.
|By An guy (Boomer) on Friday, November 02, 2001 - 06:24 pm: The Nook|
Wow- thanks Hatcher- I think I didn't read OT's oven tek as closely as I should have- I didn't do it exactly like you are saying, so I hope it worked.
But I read your explanation well enough to know exactly what to do next time...
|By Patrick (Valence) on Friday, November 02, 2001 - 08:43 pm: The Nook|
I think that removing the shrooms from the grow chamber is where the most contaminations occur. What I do is take a zip lock bag and spay the insides with very little lysol.( barely wet). Then I close the bag and rub the bag to make contact with the lysol all over the inside. I put this inside a shoe box, which has been sprayed with lysol, upside down and slightly open with the top on to let it dry completly. ( just a few minutes depending on the amount of lysol used in the first step) I then close the bag untill the cap is ready to spore. I open the bag inside the grow chamber just enough to get the cap in the bag. I then rip the stem in half where the bag opening is. Then close the bag. The mositure from the shroom rewets the lysol and it kills ( maybe not but haven't had any contamination yet)any errant spores that might be on it. I then pull off the stem while still in the bag. Then into the oven where I open the bag and plop it down onto something ( have used paper and glass ) cover with a clean jar and wait. Then its standard oven spore tek. I also like to take the cap after it has left some nice black marks and put it into the sterial jar. Squirt some water into the gills and draw it back out again. Makes some nice black murky water.
|By Nan (Nanook) on Friday, November 02, 2001 - 09:55 pm: The Nook|
A very good word of advice: Lysol is extremely toxic to shrooms. So is Rubbing Alcohol. Even the fumes are sufficient to kill the gills... And any effort to obtain a decent print. Lysol residue is toxic to shrooms. Lysol has a purpose, but not in Spore Printing. If you use Lysol, make sure everything is completely dried and ventilated before you begin working.
Alcohol wipes of glassware, instruments, ect. are OK, but make sure no wet surfaces or fumes come into contact with mycelia. Even the fumes kill.
This is one reason I like a cheap pressure box... You get excellent sterility _and_ working environment. Sealed jars, foil wrapped instruments, plates, etc., are placed in the box and wetted with Lysol. The filtered air from the fans evaps off the sanitizer spray (20-30 mins which also gives the disinfectant time to work), and when you introduce live myceila for printing or cloning you have a chamber of cool clean filtered air to work in.
If you use a sanitizer solution around or on live myceila (as perhaps on a mushroom cap for printing) I suggest a very weak Betadine solution in sterile water. Keep the alcohol and disinfectant spray as far away as possible.
Plucking caps with a plastic baggie is a very good idea.
|By packman (Packman) on Friday, November 02, 2001 - 10:18 pm: The Nook|
Newby first time, first time success at making an eddible mushroom spore syringe:
Made lid ala PF tek - one inoculation hole on the edge and one large 1/2 inch hole in the middle. Lid placed on upsidedown on jar.
Baked 1/2 pint jar (w/lid) in oven at 250 for 15 minutes covered with al foil.
With the jar still on the oven rack, remove the al foil and place masking tape across both holes and down both sides of the rim.
Allowed to cool 12 hours before mushroom was ready for print.
Spring water was boiled in a "boily pot" (32 oz capacity). The water was repeaditly pulled in and out of the syringe. The water was dumped, replaced, and 12ml of boiling water were pulled into the syringe. The cap was placed on, the whole thing wrapped in AL foil, and placed setting upright in the "boily pot", boiling for 15 min. The syringe was not submerged, only half way in the water. Allowed to cool 12 hours before print was ready.
The oven was heated to 300 while prepairing the cap. A 3-penny nail, philips screwdriver, and pearing knife were sterilized with rubbing alchol on a cloth. The nail was placed in the head of an eddible mushroom that had a dusting of spores on its veil. The pearing knife was used to hack through the stem as close to the cap as possible (within 1/4 to 1/2 inch).
The cap was carried into the kitchen on the 3-penny nail. THe oven was opened, the prepaired jar placed on the rack, and the lid removed. The cap was tossed into the jar nail and all. The lid was placed on tight.
2 hours later, the oven was reheated, next the jar placed in the rack, and the lid was removed. The Cap was removed quickly (but clumsiliy (touched the inside of the jar several times getting the cap out)) and the lid was replaced. The sterile syringe or water was inserted through the inoculation hole. It was squirted in and out several times. Concerned that the spores were not in the water, the screwdriver was placed through the larger hole and the spore print scraped around. The water was drawn back into the syringe, and immeadity injected into modifyed PF style jars (for my favorite eddible). Germination was evident in 3 days.
Thats what I did, quick, easy, lazy. Rubbing alchol makes sterilization of tools a snap. The cap only sat for a coupple of hours, if that long. The cap was massicered by the 3-penny nail and pearing knife, but everything worked nevertheless. No jars have shown any contaminents from this syringe.
|By Nan (Nanook) on Friday, November 02, 2001 - 10:57 pm: The Nook|
Good practical Tek
|By Hatcher (Hatcher) on Saturday, November 03, 2001 - 09:38 am: The Nook|
So Nan, ya put everything in the box, then put the lid on and force filtered air thru?
Source of said filtered air? Mainly just curious. Living in a one bedroom apt., the oven provides not only an effective route, but a viable one as well. And who know's, might get a trailer with a basement someday!
|By Nan (Nanook) on Saturday, November 03, 2001 - 10:47 am: The Nook|
Open front. Most of the time I don't bother opening the rear "hatch". Since it is pressurized.
Scroll down Archives->Gloveboxes
The box itself is based on the 9er Tek Box, with the addition of a second fan, no trash bag gloves. I flipped the container over and cut holes in the bottom... No lid to purchase (don't need a custom lid). It is a $15.00, open front, pressurized box.
The fan and filter details are down there
My box is kind of a cross between a true Sealed Glovebox and a Flow Hood of sorts.
|By Hatcher (Hatcher) on Saturday, November 03, 2001 - 06:01 pm: The Nook|
Appreciate the feedback Nan. I'm sure a proprly constructed glovebox is a good thing, but the space limitations of an apt. dweller kinda make it outta the question. The oven gets it done, then looks normal again. Later man!
|By Nan (Nanook) on Saturday, November 03, 2001 - 06:40 pm: The Nook|
Can't argue if it works for ya