|By Nan (Nanook) on Monday, October 22, 2001 - 12:30 am: The Nook|
Tapered wide mouth half pint jars
Plastic wide mouth jar lids
Camera lens paper
Sterile syringe with needle
1. Trim a piece of lens cleaning paper to a disk roughly the size of the bottom of a jar and place it in the bottom of jar.
2. _Loosely_ cap jar, then wrap the whole jar in aluminum foil.
3. Autoclave at 15 PSI for 20 minutes. Allow jar to cool to room temperature before moving on.
4. Prepare the spore collection "chamber" by cleaning thoroughly (soap and water at least) a glass baking pan and the top half of a 2 liter soda bottle that's been cut in half. The cap should be on the half bottle. Get all the supplies near the terrarium and make sure you're as washed up and sterile as possible.
5. Select a mushroom with a good cap (flat or even just starting to invert) growing in a suitably sterile environment. Go ahead and remove the jar with the lens paper in the bottom from the aluminum foil.
6. Harvest the mushroom and with a sterilised knife or razor cut the stem off as close to the cap as possible. Then, opening the jar as little as is humanly possible, transfer the cap, gills down, into the jar. _Loosely_ cap again and transfer to the pan and cover with the half bottle (the half bottle is used like a bell jar).
ASCII diagram of setup which I'm sure looks horrible in html :
7. Allow setup to remain somewhere cool and relatively draft free for a day or so until sufficient spores have been dropped
8. Remove jar from setup, quickly remove mushroom cap with sterilised forceps, pour in sterile, distilled water. Tighten the jar lid and shake vigorously. The lens paper allows you to get just about all the spores into solution without spending all that time scraping which opens up opportunities for airborne spores to get into your container.
9. Open jar as little as possible again and use freshly sterilised forceps to remove the lens paper. Recap tightly.
10. Make a spore syringe initially by heating the needle red hot and piercing through the jar lid. Suck up the spore solution and remove and cap the syringe. Seal the hole with good quality plastic based tape and store in a light proof container in the fridge, removing the tape and loading syringes as needed. You can re-use lids by taping the hole after a jar is emptied.
The beauty of the this method is that the existence of the one piece cap turns the jar into the equivalent of a culture flask or dish in the lab: the loose cap allows gas exchange but still prevents *almost* any contaminant because of the nature of the air flow. The idea of the soda bottle is great because it's easily obtainable, still allows gas exchange, helps keep humidity high, and contributes to sterility.
Lens paper is very fine, thin, and porous. When it comes time to make spore water, you only have to open the jar long enough to add sterile water then reseal. Shake well and get instant spore solution, schwing!, no need for risky mucking about to scrape the print. You don't even need to remove the paper if you are going to load all your syringes immediately, you just don't want to let the cellulose of the paper become a substrate for the spores to germinate in solution on. The caps are cheap enought (about 25 cents a pop) that they're disposable if you don't want to reuse them.
originally posted by 'spore monkey' - See: Mal's Print/Syringe Tek
|By Nan (Nanook) on Monday, October 22, 2001 - 12:34 am: The Nook|
Multiple Spore Printing Tek
This is an improvised version of a popular spore printing idea that already exists. The concept is to allow a mushroom cap to drop its spores on a sterile medium, producing a viable "spore print". The cap must be covered to protect contamination by airborne mould spores and to keep the humidity high so that the mushroom will drop its spores. There are many sterile mediums available but I would recommend aluminium foil above all of them. Paper does not typically allow spores to adhere properly, making it unsuitable for mailing. Also, it is harder to seal without leaving it prone to contamination. The bonus in using this method is the ability to produce multiple sterile/viable spore prints.
Baking Tray (no/slightly raised edges if possible)
Pyrex Baking Dish (upside down it must fit entirely within the tray)
Hand Towel/Paper Towel
Alcohol Aerosol Spray (glen20 is quite a common brand)
Sharp Blade (cheap retractable pen knives are available at most supermarkets)
1. The spore printing "setup" needs to be constructed. This is really quite simple. Take the tray, lay down enough towelling to cover the whole tray, turn the pyrex baking dish upside down and place it in the centre.
2. Now the actual printing squares have to be cut out. You want each individual square of foil to be slightly wider than the cap you want to print from, and twice as long as it is wide. Cut out as many rectangles as will fit in the setup, or as will be printed. Lift the pyrex baking dish and neatly place the foil squares underneath. (I prefold them in half, then unfold them before I put them down. This will make more sense later on.)
3. Place the entire setup in the oven at 150C for 30-60 minutes. All materials used (with the exception of the towelling) are good conductors of heat so 150C is plenty hot enough to sterilise the setup. 30 minutes is fairly short, yet I have had no problems with contamination using this time period. Be sure not too sterilise with too high a temperature for too long or the towelling may ignite
4. Remove the setup from the oven and while (relatively) hot create a "handle" from the tape. I find this necessary because unlike small, single print covers a curved edge pyrex baking dish is quite awkward to lift with minimal contact. Cut a measure of tape, fold it in half, but not completely! Only fold it to about halfway down the length so that you end up with a sort of "T" shaped piece of tape. The top of this "T" should be adhesive whilst the sides are not. Stick one sticky side on the top of the pyrex baking dish, and the other on the side of the pyrex baking dish. If you can not understand this handle concept (I know it is not very clear!) then feel free to improvise your own. The reason this is done while hot is to ensure a more permanent contact between the tape and the dish. If it is done later the tape often becomes unstuck after being sprayed with aerosol alcohol.
5. So now you have a nice sterile setup! Well, except for the tape of course! So leave it somewhere relatively clean to cool. Room temperature is what you are aiming for here. You will find that the pyrex baking dish will take quite a while to emit all heat and return to room temperature (approximately 2 hours).
6. Now take your setup and place it somewhere partially enclosed to minimise chances of contamination by airborne particles. Bring your terrarium closeby if possible, this is where a nice portable terrarium comes in handy. (If one is not available you might consider making a mini-terrarium for transporting casing/cake to allow printing, or alternatively, place your casing/cake in a clean plastic container. Thoroughly spray down the surrounding area with aerosol alcohol. Don your dust mask and gloves. Spray the gloves for extra sterility. Wipe down the sharp blade with an alcohol wipe (make your own by spraying a tissue) and hold it in the enclosed area while the alcohol evaporates. Give the immediate area another liberal spraying (can't hurt to be overcautious
7. Now you have a relatively sterile area. Aseptic is quite a difficult goal for the avid home cultivar, so relatively sterile will have to do (in my experience it works fine most of the time anyway). Remove the lid from the terrarium, select the choicest specimens (any that developed larger, faster, or generally more roust than the others are desirable). The veil should have released itself, leaving it drooping around the stem. The cap should be hemispherical, approaching a slight flatness. While this will result in a somewhat smaller print than an upturned cap it means that only the rim of the cap actually touches the printing medium, thereby lowering chances of contamination.
8. Now the cap must be cut from the mushrooms. Using the ethanol sterilised blade make a cut at rought the same stem height as the edge of the cap. It is best to cut as close to the gills as possible, but it is also best not to touch the gills with anything at all if possible. I figure edge of cap height is a good compromise. Some people sterilise the cap by wiping the top down with a hydrogen peroxide soaked cotton wool ball, then the same with ethanol. Some people even *LIGHTLY* wipe the gills. I have never found any of these necessary and I advise that they are not attempted unless contamination has become an issue.
9. Slowly lift the dish using the tape "handle". Slow is the key here because a quick lift will result in a large influx of air, raising chances of contamination. Once lifted to a minimum height (enough to allow a gloved hand to fit in) place the cap in the centre of one side of the foil rectangle. Once the process is completed for all caps the setup must sit for 12-48 hours undisturbed. The longer you leave it the heavier the print. This is, of course, just a rule of thumb and mileage will vary between individuals. Each individual microscopic spore takes between 30-60 seconds to drop, so do the calculations and you can work out the time needed for a nice visibly thick print
10. After waiting 12-48 hours, spray down the entire area with aerosol alcohol again, put on the dust mask and gloves, and spray the gloves again. Have your baggies nearby and open the first one. Slowly lift the dish, remove the cap carefully and quickly fold the foil over. Now fold each side over slightly, and finally, fold the end. Place the print in the ziplock baggy and you have just created a spore print. Repeat the process with each cap and you will have a nice batch of prints. Some people suggest removing the caps with a sterilised pin, but again I have never found this necessary. Plus, the outside of the print has to be touched to be folded anyway. By lifting the cap carefully your hands should not come into contact with the printing surface.
11. If you regularly make batches of prints like this for the purpose of distribution (be it sale, trade, competition etc.) a randomly selected print should be made into a syringe as a sort of "quality testing procedure". Label your prints with small adhesive labels that include species, race, printing time, print date, and source. This will be appreciated by all who receive your prints. Good luck and happy printing!
originally posted by "auto59009"
|By Nan (Nanook) on Monday, October 22, 2001 - 12:42 am: The Nook|
I use a simple method that's worked well for me. I use aluminum foil. I cut a 6" wide strip off the end of a roll, then fold it over in half. That give you about 3" by 12". I then put a cap every 3-4 inches, depending on the size. Then, the top lays over the caps, and the whole thing is stored in a small airtight box for 24 hours. Remove the caps, cut the foil into individual prints, then fold the edge of the foil over making a seal all around the print. Use a "sharpie" to label the print. Minimal work, and if the caps are from cultivated fruits and clean the prints come out clean. Foil is nice bacause you can scrape all the spores off it, I prefer it to paper, although wax paper is ok, but you have to tape the edges, whereas foil can be "self-sealing".
|By Nan (Nanook) on Monday, October 22, 2001 - 04:23 pm: The Nook|
SPORE PRINTING EQUIPMENT
KERR 1/2 PINT WIDE MOUTH (LOW FORM) CANNING JAR. (ANY SUITABLE JAR IS OK)
FINGER NAIL CUTICLE SCISSORS - (cosmetics - drug stores)
ALCOHOL, TEQUILA SHOT GLASS AND EYE DROPPER.
1. Presterilize the jar and regular metal lid (rubber edge up) in a small toaster oven at around 300 degrees Fahrenheit for around a half hour. Keep the lid loose during the sterilization cycle. When the jar has cooled down, tighten the lid until it is time to use the jar for a spore print. The rubberized edge will be a bit melted, but that won't be any problem in this technique.
Note: What follows is a sterile technique. The first rule that must be always followed is to wash hands prior to sterile work. Hands are a prime source for bacteria and microspore contaminants. Sterilize all the work surfaces with rubbing alcohol. Minimize drafts. Try for a still air environment. Don't breathe on the work. Run a small home appliance style HEPA air cleaner (99.97% rated efficiency - available at drug and department stores) for a few hours in a closed room to clean the air before doing sterile work.
2. Flame sterilize the scissors with an alcohol flame and snip off the mushroom cap. Cut the top of the stem as far up into the cap as possible so that the gills of the mushroom will sit flat on the surface of the jar bottom. With quick and sure movements, place the cap into the jar and place the lid on loosely. Pierce the top of the cap with a straight pin to pick it up and handle it.
3. Leave the jar with a loose cap for a couple of days in a draft free area away from direct sunlight. After the print is taken, quickly and with as little air disturbance as possible, remove the jar cap and extract the mushroom cap from the jar. With a loose jar cap, let the jar sit in a draft free place to dehumidify for a few days before sealing it up (with tape) because there will be some residual moisture left behind on the spores and glass. Store the spore print jar at room temperatures in a dark place away from sunlight. Don't store it in a refrigerator.
Psilocybe Cubensis spores begin to degrade a few months after they are taken. After approximately 1 1/2 years, spore germination will be greatly reduced or won't occur at all. Germination is massive and quick when the spores are fresh.
MAKING A SPORE PRINT Unknown
A spore print is a collection of spores on a flat surface. It can serve several purposes. It can be used to assist identification of the specimen by observing its color or if made on a glass slide, by studying the shapes of the spores under a microscope. Mycological identification keys include descriptions of spore prints and microscopic spore features for different species. Spore prints are also the standard method of collecting spores for later germination on agar media. A print from a single mushroom cap contains millions of spores. Many mushroom lovers are now making spore prints on paper from species available in their locales and mailing them to cultivators in other areas where such species are not found. Secret spore exchange correspondence clubs are becoming quite the vogue and will probably be more common in the very near future. A word of caution regarding this practice should be given, however. Do not assume that spores received in this manner are from the species the sender claims they are. If the sender has misidentified the specimen and the recipient cultivates and ingests mycelia or extractions therefrom, the result may be disasterous. Furthemore, I would not put it past some anti-drug fanatic to purposefully disseminate spore prints of dangerous mushrooms to amateur cultivators. This could result in sickness and death for thousands of persons.
To make a spore print take a mushroom with it's cap fully opened and gills exposed. With a sharp sterilized blade cut off the stem as close to the gills a possible. Place the cap gills-down on a clean, white sheet of paper, or on a sheet of glass that has just been swabbed with alcohol, or on two or four sterilized microscopic glass slides. Cover the cap with a clean, inverted bowl or bell jar to prevent drying of the cap and intrusion of foreign organisms. Let this stand as such for 24 hours. If a good spore print has not been formed after this time, tap the cap lightly with the flat side of a knife or spatula. This should shake loose many spores. If the print is made on glass, cover it with another glass sheet immediately after removing the cap to prevent contamination. If microscopic slides are used, place two face to face and seal the edges with tape. If paper is used. fold it several times so that the print is well inside.
"MAKING A SPOREPRINT" - Shroomwizard Print Tek
I believe everyone who cultivates mushrooms should make sporeprints, for their own private use in growing future crops and also to pass onto others who are willing to try their hand at the growing process.
1.) Take a clean washcloth and spread it out in a place it will not be disturbed for 24 hours, I just put mine on my dresser top.
2.) Now take a 3 x 5 index card and put it on a cookie sheet in an oven for 10 minutes that has been preheated to 200 degrees.
3.) Remove the index card from the oven and let it cool down for a few minutes.
4.) Take the index card and lie it on top of the washcloth.
5.) Take a "fresh" mushroom that has a fully opened cap and cut the stem off of it as close to the cap as possible.
6.) Lie the cap, gills facing down, on top of one half of the index card and cover with an inverted bowl or cup.
7.) Leave this alone for 24 hours (48 hours if you want an excellent print). As the mushroom cap dries it will drop its spores onto the index card.
8.) When the waiting time is over you can remove the inverted bowl or cup and slowly pick up the mushroom cap. What you should see is a beautiful sporeprint on the index card where the mushroom cap was; I like to think of this as God's fingerprint because it resembles one.
9.) Fold the index card in half, enclosing the sporeprint, and put tape around the edges to seal. This will prevent any foreign spores from entering your print and also prevent any of your spores from getting out.
At this time I should mention that I place my shroom cap on one side of the index card so that when I fold it in half I won't be folding the actual sporeprint itself. A print made this way will last for many years without any refrigeration, I place mine between the pages of a book, that way only I know the best books to read (he-he).
A lot of people write to me and ask me the purpose of the washcloth. It is so that a small amount of air can seep under the bowl and help draw the humidity out of the mushroom cap. It is the moisture in the fresh mushroom cap that the spores are stuck to, and as the shroom cap dries out the spores will be released and fall on the index card. After you make your sporeprint, if the mushroom cap still looks fresh and the sporeprint you made is very dark in color, you can place the same mushroom cap on another sterilized index card and leave it there for 48 more hours (using the same process) and hopefully get another dark sporeprint from it.
Gadget Guru is here, I have an idea for a combination glove-box flow-hood, that is top of the line. The spore print procedure is this: I took If I took a fish-hook and installed it on the top of the jar-lid. From the hardware store get small Stanless steel bolts and washers. You will have to bend fish hook and drill hole in center of jar lid. Assembly required. Have say 15 or so made, Put some sterilized water in jars and steam sterilize, store them in glove box. When caps curl hang on hook (There should be water in jar, a little bit.) When cap has deposited in water Have good syringe with sterile water inside, you Know Boil water 20 minutes suck into syringe a couple times, Then suck up and leave water inside and wrap wih aluminum foil put in pressure cooker (You can use cooker with fish hook jars, put them all in the same pot. Steam sterilize store in glove box. Now that you have it all, Remove foil from syringe, shoot water from syringe into jar, oh remove cap and ingest. Do the Martini stir and fill syringe, repeat, by storing in glove box say 3-5 syringes you wont run out. And one of the best features is that you do not ever have to buy disposable again. and another one of the features the syringe has is a control on top of syringe, it will stop at 1 cc. I wil shoot 1 cc into each hole squeeze and go to next hole. I have lamp for steralizing and i can reach it from inside box. Well more LATER
|By Kevin Smith (Canshroom) on Wednesday, October 17, 2001 - 09:55 pm: The Nook|
I've heard that getting viable spore prints is harder than it might seem. How difficult is it really? Also, are there alternative methods to perpetuate further generations of mushrooms other than by using prints or buying prints or syringes? Thanks.
|By Nan (Nanook) on Wednesday, October 17, 2001 - 10:25 pm: The Nook|
Good questions. I am going to send you down to the Archives for some material on this. It's a good place to start on your answers.
The first topic you want is: Archives->Spores
Also check out: Archives->Honey/Dextrose Q&A and Archives->Cloning
Keep us posted.
|By Marx2k (Marx2k) on Saturday, October 27, 2001 - 08:42 pm: The Nook|
The best technique I have for spore printing with means of sucking it into a syringe is taking a half pint jar half full of sterile water. I then cover the top of the jar with tin foil... cut a hole in the tin foil large enough to rest the cap on without it falling through. I then rest the cap on the tin foil. I then cover the jar with the lid/band. After a day or three, there is a heavy heavy spore print in the water. I quickly remove the cap/tin foil, re-cover the jar... shake up the water and then draw syringe after syringe of perfectly sterile spores.
Easiest way to do it for a syringe!
|By Marx2k (Marx2k) on Saturday, October 27, 2001 - 08:46 pm: The Nook|
And the easiest way of getting sterile water and the foil already there without much work? Fill the jar halfway with sterile water. Place the tinfoil, cut a hole, cover with band and lid. Drop that bitch into the pressure cooker for hoever long you want (Anything more than 30 minutes is fine). When it comes out and cools, everything inside is sterile. In front of the oven, remove the band and lid and put the cap on, recover band and lid and follow above instructions. When taking cap out, grab the tinfoil and take the cap out with the tinfoil. What youre left with is just spore water. All sterile and nice. Make sure to have a hold in the lid (covered with tape the whole time) to suck up the water through into the syringe so you dont have to open the jar afterwards.
|By Nan (Nanook) on Saturday, October 27, 2001 - 08:52 pm: The Nook|
Good tips, destined for the archives.
|By Nuecrew (Nuecrew) on Wednesday, October 17, 2001 - 03:43 am: The Nook|
I bought a spore print sent to me in a zip lock little bag. What if I cleaned the outside of the baggie, inject the baggie with sterile water and suck the spore water back out. Are spore prints sterile as sent from spore suppliers?
|By Nan (Nanook) on Wednesday, October 17, 2001 - 04:05 am: The Nook|
That's a good Tek with the plastic bag, use it. All suppliers state their spores are sterile, it's not always true (tho I have _never_ heard of a contamed syringe from PF.) Good Luck
|By Trippysmurf (Trippysmurf) on Monday, October 22, 2001 - 06:42 pm: The Nook|
i made a print from a cap that i put in a new baggie, i put a pin in the middle of the cap to minimize the plastic touching the cap (didn't work too good), there was some moisture on the top part of the plastic when i took the cap out, i found a very dark thick print, but it was wet, is this print still good? i plan to do the injecting water into the baggie too, but i think ill wait till the print dries, so is the print still good?
|By SYDYSTYK (Addict) on Monday, October 22, 2001 - 06:45 pm: The Nook|
try the honey tek to see if it was good, if it grows mold/contams you will know
|By Nan (Nanook) on Monday, October 22, 2001 - 06:59 pm: The Nook|
Trippy, you are going to have to test it to find out. We have no way of knowing. The print should be dried if you intend to store it for any length of time, open the baggie and lay it in the Desiccator and get the water out.
Liquid Culture is an easy, cheap way to test for germination and contams.
|By The Roc (Rochester) on Monday, October 22, 2001 - 10:57 pm: The Nook|
I placed a cap in a baggie and let it print then injected water and made 3 syringes... worked great for me. I have used 2 of the 3 and am very pleased with this method.
|By Marx2k (Marx2k) on Saturday, October 27, 2001 - 06:37 pm: The Nook|
The Roc, does the cap touching the baggie not have a problem with sterility? I thought that the cap should not touch the medium you're printing to?
|By Liberty_Caps (Liberty_Caps) on Saturday, October 27, 2001 - 08:32 pm: The Nook|
I was wondering,, I'm makeing a hawiian print right now, and have been to PF's site and it says I need a glass stiring rod to scrape the spores into the water. but I do not have a glasse stiring rod. do I really need one of these?????? or could I just shake the jar to get the spores and water mixed????? probley not,, so what do u guys use to get the spores evenly distributed in the water????? should I just shoot the steril water onto the print so the water pressur will loosen the spores up,, then suck the spore water up into the syringe and repeat the prosses a few times????????
I don't want to screw this up and I don't want to have to take off the lid to mix the spores up
so any advide would be greatly appreciated thanx!!
|By Marx2k (Marx2k) on Saturday, October 27, 2001 - 08:37 pm: The Nook|
You can use anythign sterile to scrape the spores into the water. You can use the sterilized needle of the syringe youre going to be sucking the spores into. You can even use your finger provided you flame sterilize and alcohol wipe it
|By Marx2k (Marx2k) on Saturday, October 27, 2001 - 08:37 pm: The Nook|
Ohm ans as long as the water is sterile, hell... you can just shake the jar up and this will mix up the spores nicely.
|By Nan (Nanook) on Saturday, October 27, 2001 - 08:55 pm: The Nook|
Flame the tip of an old dull knife, it works for me.
Good to see you back Mr. Caps
|By The Roc (Rochester) on Sunday, October 28, 2001 - 01:23 am: The Nook|
Marx2k I just propped up the opening in the baggie with a toothpick and allowed some airflow and had no contams related to the cap touching the bag. I was more concerned about a release of moisture during the print...
|By Liberty_Caps (Liberty_Caps) on Sunday, October 28, 2001 - 03:47 am: The Nook|
alright then thanx alot guys.
I just checked my print and it's going great I can see lots of purple spores collecting on the bottom of the jar,, so in a few more days I'll be ready to make my syringe
and I also got a flat cake colinizing right now my first attemt and it's looking good so far
|By Nan (Nanook) on Sunday, October 28, 2001 - 04:09 am: The Nook|
|By Liberty_Caps (Liberty_Caps) on Sunday, October 28, 2001 - 05:42 am: The Nook|
sorry but I have one more quiestion,
if I'm makeing a print and after the mushroom cap dropes its spores, do I have to let the print sit in the jar for a few more days to DRY or can I just suck up the FRESH/NOT DRIED spores into a syringe
I heard fresh spores were better but don't really know so any help would be GREAT!! THANX
|By Nan (Nanook) on Sunday, October 28, 2001 - 08:41 am: The Nook|
Yeah, if you intend to use the spores right off you don't need to wait or dry them. Go ahead and load syringes and shoot as soon you are ready.
|By Liberty_Caps (Liberty_Caps) on Monday, October 29, 2001 - 04:05 am: The Nook|
alright thanx, ok so I just made my syringe and I scarped the spores into the water then sucked up the print,, I only made 1 syringe out of the mushroom cap and well,,, my syringe is BLACK I noticed when I took the cap out of the jar the print was really dark purple,, so maby I just have a shit load of spores in this syringe or I don't know what???? any clarification would be great thanx
|By Nan (Nanook) on Monday, October 29, 2001 - 04:31 am: The Nook|
Just a lot of spores
|By jim brown (Shrhobbyist) on Monday, October 29, 2001 - 08:56 am: The Nook|
That is a lot of spores. I suggest you either dillute it to make more syringes or use very little (.25cc) on each jar. Otherwise it would be kind of a waste of all those spores on a dozen jars when you could get a lot more out of it. Good luck, let us know how it works out, it's great to be self-sufficent spore-wise.
|By Kevin Smith (Canshroom) on Sunday, January 20, 2002 - 07:01 am: The Nook|
After all the time I spent concerned about contams while printing, I had success on my first attempt. I tried the spore syringe tek in the archives and except for a few glitches, I worked. For one, the water boiled out of most of my syringes. The other problem was scraping the spores into the water. This time I'm suspending the cap over sterile water a la another tek I saw here. Anyway, despite the few glitches I ended up getting 3 syringes from one print and now every jar is being colonized at every inoculation site. Nice, cottony growth and spreading by the day. Thanks everyone for your advice.
|Posted by: goatywoaty Dec 28 02, 03:25 AM GMT|
| im having very horrid contam problems and i think this is why:
wipe w/alcohol napkin
scrape print into
steralized dish with sterile water
sy sterelized by sucking up boiling water and hold ing for 5 minutes, then h202, then alcohol.
sucked up spores
i think thats where my contams is coming, approx 5 attempts now have been various colors of mold, black, red, white, green, grey... its very frustrating. im doing everything by tried and true teks ive read for pf tek and millet... any suggestions as to minor things here and there i may not have picked up, or am i just releasing spores @ jars somehow...
|Posted by: steveoi812 Dec 28 02, 04:05 AM GMT|
|do you pressure cook your jars of substrate or boil/steam? The print could be contaminated all together. A glovebox might be a good investment. Where do you incubate your jars?|
|Posted by: Malformed Dec 28 02, 04:07 AM GMT|
| if you havnt allready, build a glovebox.
i use prepackaged sterile syringes.
i sterilize distilled water in jars with plastic lids, with foil over them.
i take the spore print inside a pre-sterilized jar, on top of a sterilized aluminum square cut from a soda can.
take a shower, brush teeth, rinse with listerine, decapitate the shroom, place it in the jar, on top of the aluminum square (the side with no paint)
once the print is taken (a day or 2 later), and all my supplys are in the cleanest room in the house, (my grow closet.)
i brush my teeth, rinse a few times with listerine, take a shower, use antibacterial soap and shampoo. (neutrogena is great)
i put on clean cloths.
i unwrap and open everything very slowly as to not create a breeze.
remove the mushroom cap from the jar, take out the print, and scrape it directly into the jar of water. it should still be a wet print, if you think youve been realy clean. just drop the whole print into the jar, put the lid on and shake it up damn good. it should make a nice dark jar of spore juice.
you should beable to make 15 syringes or so from one half pint of spore juice.
keep in mind that a single spore print could make alot more than just 15 syringes...
i just like to keep the batches small incase they are contaminated somewhere along the printing process or making the syringes.
obviously if you make 100 syringes at once, and mess up. you will be alot more pissed off than if you just made 10 - 15
once i got the syringes made, i inoculate 3 or 4 jars, with seperate syringes from that batch to test them.
out of the 4 or 5 batches of syringes ive made, only 1 batch was bad.
i do not have, or use a glove box.
but as you can tell im very clean and i still get contaminated syringes.
my recomendation is to use a glove box.
and if you are at all serious about this 'hobby'
buy a presure cooker if you dont have one!
even a cheap small one so you can sterilize a few things to make prints.
razor, something to print the cap in and on. and some water.
plus, you can wrap some used syringes up in foil and sterilize them in the presure cooker as well and not have to mess with boiling water.
|Posted by: DirtyWOP Dec 28 02, 02:41 PM GMT|
| Is this being done in open air?
If so, there is no way you could possibly do that procedure and NOT get contams....get a glovebox like malf said....
also....that is not a good way to sterilize a syringe for more than one reason. You say you suck up boiling water, then h202, then alcohol. You should have stopped at the boiling water.......alcohol will kill spores. In fact, sucking up boiling water isn't a very good method. I've used it, but 50% of the times it resulted in contamination.....
Steam sterilize your syringes in foil....fill them half with water, pull out the plunger, tape it in place, put the cap on, and steam sterilize or PC. Flame the needle red hot before each use.
Better luck next time around man....
|Posted by: Fungusmaximus Dec 28 02, 03:03 PM GMT|
| All good advice. PC, PC, PC! pRESSURE COOK EVERYTHING [email protected]
And a glovebox is also a great addition to your equipment, its a whole other ball game when you play w/gloveboxes/flowhoods etc.
Do some looking in the archives, find a design that fits your needs and build it.
*** whispering quietly***
If you build it ....contams wont come.....
|Posted by: 420M Dec 28 02, 03:24 PM GMT|
| I think I am going to build a new flowhood.
|Posted by: Malformed Dec 28 02, 05:55 PM GMT|
| oh, after gargleing listerine i also wear a facemask.
if i had a hair net i would wear that too.
and, gloves. i dont have any sterile nitrile gloves (yet) but i plan on buying a box soon as i start working with compost and pan cyans.
i also want a flowhood, but a positive presure glovebox will be fine for a while. vitti said i could have his since he doesnt grow anymore.
but a presure cooker, even a cheap one is a good investment.
i like to go 30 - 60 minutes depending on how much i have in the pc.
even though i do all my stuff in open air right now i realize its not a good idea and i do get contamination here and there.
Build a glove box
Get a pressure cooker