Anyway been alot of questions about this. I have been using this for years. here you go. in this case i use a shmuvbox. it is a hepa with a big clear trash bag over it, you place all your clean tools, fruits for cloning, and benzo torch to sterilize your blades in this bag. usually i will wipe down every single part of this hepa with 70-90% rubbing alc. be anal, spray it with disinfectant spray. this thing will pour plates and take clones easily. holmes brand hepa filters with a small ionizer built in are a great size. you only need a small room size filter. and it should be around 30$ at walmart. once you put the bag around the filter, tape it down to the table your working on. this will in turn, allow the bag to stay put. now blow your bag up that has all your cooled utensils and sterile equipment in it. turn on the hepa and when it blows up, cut two holes in it for your hands. one for each. and if you can't find 100% large lawn bags for the bag, you can cut eye holes as well. this produces positive air pressure and forces out the bad stuff. its real simple, clones can be taken and it stores nicely. its efficient and takes up little space, and hides easily. you can also just do all this work in front of a flowhood or in a glovebox or whatever you use for sterile work. sam's club has large 55 gallon clear bags. you can fit at least 20 quarts and have room to work in that size. and its about 10 bucks for 220 of them. ive just bought my 2nd box of bags after 5 years. 220 will last you clean work for close to 5 years doing at least 1 or 2 clean sessions weekly. Pointer: Use the lowest airlfow setting of your hepa unit. Credit goes to Molestor from the Nook. First take a shower. Then you can clean your work area and bring in all utensils and supplies prior to starting all of this. I turned off the air condition 30 minutes prior, I wiped down the table with bleach wipes, rubbing alcohol and sprayed lysol everywhere. The lysol was sprayed in the air, on the table ect. 1. Take your hepa unit out of the box. 2. Open up the housing that holds the filter. The first time you insert the filter. This example is a 5 year old filter. 3. Get a paper towel and douse it with rubbing alcohol. Completely douse it. The more the merrier. I use 90% isopropyl. 4. Begin to wipe all parts inside this area where the filter sits. 5. Hose down the inside with lysol. You only need to hit the areas that are exposed inside the filter area and the parts on the hepa unit that will be exposed inside the glovebox. You will see in a later step. 6. Make sure to also wipe all plastic parts on the housing for the filter itself. The black plastic part you see in the picture. Lysol the plastic as well. 7. Set the filter back in the housing and place the black filter material that comes with it back over the filter as shown. 8. Do the same thing for the filter door. Rub it drenched with alcohol. Then liberally spray it with lysol all over the inside and the outside. Also be sure not to set any parts that will be exposed down on the table or don't let them come in contact with anything. I am only doing this in the pictures so that i could take the picture. I then re-wiped and sprayed after the picture. You could also use bleach wipes or a bleach solution for the alcohol part if it makes you more comfortable. 9. Spray the outside of the face where the intake will be with lysol. 10. Now do the same thing(bleach or alcohol and lysol) for all parts of the shmuvbox that will inside your clean work area aka the 55 gallon lawn bag. 11. I tend to steer away from using bleach inside the working part area. 12. Take your 55 gallon trash bag and unfold it two times so that it is fully expanded but the two sides are still stuck together. bag laid out on table first unfold 2nd unfold 13.Open up the bag just enough so that the unit fits into the bag. 14. set your shmuvbox in the bag. Making sure that all the areas you rubbed down with alcohol are the only areas that will touch the inside of the bag. take note, your hepa unit should still be wet with rubbing alcohol as your doing this. 15. Cover the unit with the bag. 16. Tape down the side of your shmuvbox so that it is secure to the side of the hepa unit. 17. Turn on your hepa ionizer and the fan on to the lowest setting.
EZ Shmuvbox Setup: Setting Up A Shmuvbox
18. Let the bag fill up until it is full.
19. Now take your scissors that you have cleaned with alc or bleach wipes and cut 3 holes. One for your eyes, two for your hands. make sure that if your cutting the eye hole it is big enough so that the biggest thing you need to place in the bag will fit through the hole. i generally make the eye hole a little bigger. sometimes i have to move at angles and things to see if the eye hole is too small.
20. Now begin to wipe down all your instruments with bleach wipes or rubbing alcohol prior to placing them in the bag. If you have pressure cooked your utensils, take them directly out of the pressure cooker and place them in the bag. i generally wrap all my scalpels, spore loops and material in foil. so the inside of that foil is sterile. if your worried douse the outside of the foil with lysol before you put it in.
21. Load your jars and utensils into the bag through one of the holes you cut for your eyes or hands. You have the option of wiping down each jar with rubbing alcohol or a bleach wipe prior to placing it in the shmuv. ive tried spraying lysol on them but i find this is to my detriment. with gloves on this can be very slippery and difficult to work with. the alcohol on the other hand will evaporate off by the time your ready to do work. you can easily spray more lysol in the bag. If your taking your grain jars straight out of the pressure cooker and quickly placing them into the shmuv it will work fine. but you can never be too careful so it won't hurt to wipe down each jar all over with rubbing alcohol. ill then shake it inside to redistribute moisture in the jar.
here are pictures of what the jars look like pre shake and post shake.
22. Loosen your jar lids in the bag.
23. You can see inside the shmuvbox now it is almost ready to use.
24. I then spray lysol in the eye hole. I wait about 5 minutes for the air to flow a little. Then clean work is ready to begin.You can use OB gloves for horses for arm length gloves or just clean your hands with alc and warm soapy water. I took a shower prior to doing all of this.
This is a picture of the glove out last time i used the glovebox.
25. Now your ready to G2G transfer or P2G transfer. You can also do any clean work you need to do like clones, liquid inoculants, spore syringes or agar plates. you can see inside how much room there is to work in, its very spacious using 55 gallon bags. that is 10 quarts but you can easily fit 20 quarts in one bag sitting. Also, a pointer is to keep the work area near the front of the bag closer to your hands. not right in front of the holes but close enough to reach. ill then place all of my quart jars off to one side. what i do is once the jar is spawned, i will push it to the opposite side of the bag so i know that the jars are complete. by the end all jars are on the opposite side that you started on. you can see this in the picture. i left it as is after the transfer to show that all the jars on the left have been pushed over after transfer. the last jar transfered was on the right and then it would be pushed to the left. this is just personal preference so i can keep track of which jars were spawned. i also wipe my and hands down up to the elbow prior to placing them back in the bag anytime i may take them out. if your using ob gloves then make sure to wipe them down with rubbing alc prior to putting them back in. I will even go the route of spraying lysol again.
G2G and P2G Using A Shmuvbox
this is just for those of you who are interested in.
this is a master PF 1/2 pint jar. it is scraped with a clean fork ala g2g to grain jars. the grain jars were prepped as per Fooman's Grain tek
here is a shot of the shmuv. this shows all 3 holes. you can see the bag blown up with positive clean air pressure from the HEPA with ionizer.
this is a view into the shmuv. you can see the grain jars and the PF jars awaiting transfer. its real simple. get fork, scratch PF cake with fork. quickly open up a quart jar that already has lids loosened. then just dump the mycelia into it. scrape again with fork then dump into lid. make sure you close lids real quick. quicker the better. its ok for the PF jar of mycelia to be open air. also, if you flip the inside of your PF lid upside down this also works as a fail safe way to rest your fork in between transfers. just set the clean part of the fork your scraping with directly on the inside of the lid and make sure it doesn't touch anything. with practice, u will have it down speedy.(this method is in the Vaults and works well)
this is a side shot of the tape holding the bag on. this is for clarity, i tried to explain it really well but closeups always help the view..
this is the view of the bag over the top of the HEPA . right where the buttons are.
this is some random black hand(quite possibly snake eyes from gi joe) scraping the PF mycelia with a fork. you can easily use 1/2 pint mycelia to knock up 10-15 quarts. and as you can see, the bottom of the shmuv is used as a garbage can. just scrape off the verm barrier right into it. this is thrown away or used as a trash bag afterwards.
this is after the mycelia is dumped into the grain jar
this is a half ass scrape.you really want to make it almost powdery. that way you have more inoc points. it colonizes faster. generally u can have 100% colonized jars this way in 5-7 days. the more mycelia the less time. so if you put alot, you can get full colonization in 3 days. just up the ratio.
after that shake the jar. let it colonize 3 days then shake again. you should have fully colonized jars from g2g in under a week.
2014 Update
Here is a sequence of growth in P2G(PF to grain transfer)
The strain is APE(Albino Penis Envy) and the spores are over 5 years old. All transfers done with the above method.
Inoculation 8-6-14
Day 1: 8-7-14 Incubation
Day 2: 8-8-14
Day 3: 8-9-14
Before a Shake
After a shake
Day 4: 8-10-14 A day after a shake
Day 5 8-11-14
Day 6: 8-12-14
It looks like it will be under 7 days after inoculation full colonization using a SHMUVBOX!
Day 7: 8-13-14
STEM TISSUE LIQUID CLONING
These are old pictures showing a stem clone with vacutainers.
the black plastic ring is the attachment for the base of the oster blender that you screw onto the metal blade assembly then blend. you can see some vaccutainers and two syringes and one fat pin. it was solid.
This basically is the methods i use with adaptivities to it. It incorporates 9er tek cloning method, Slurry in a Hurry Tek, and some simple things to preserve your clone material for later use in a small fridge.
Supplies
- vacutainers( if you wish to store the stem tissue in them)
- as many sterile 10 cc syringes or 60cc syringes as you want clone material
- bernzomatic torch
- 1 scalpal
- 1 exacto knifes
- osterizer blender, black base, metal blade assembly, gasket ring
- regular mouth 1/2 pint jar
- distilled water
- alcohol swipes
- foil
1) First of all, I start this entire cloning process using a shmuvbox as my glovebox. You can use a SAB or Flowhood.
2) This is basically 9er tek Im just posting a walk through. You can use an oster blender base with the blade assembly and regular mouth 1/2 pint, pint, and quart jar attachements depending on volume of liquid inoculant you want. Fill a 1/2 pint about halfway with water. a metal band will easily screw onto the top. you then put foil over it. also wrap your scalpal and exacto blade in foil. pressure cook for 20 mins at 15psi. Allow to cool.(exacto blades are easy to replace and a 50 pack of replacement blades will last a lifetime.)
http://www.shroomery.org/8433/The-9er-Tek
3) First wipe down your bernzo torch, your black blender base with bleach wipes. Bring in all your sterile syringes into your clean work area.
4) Start to connect all of your syringes together. Make sure to open them quickly and quickly screw on each needle. I use the sheath as a means of putting the syringe back into it before i do all my syringe prep.
5) This is a shot of all your supplies ready to go.
6) Quickly and cleanly bring in your clone specimen. In this picture you see APE(Albino Penis Envy)
7) Turn your bernzo torch on. It has a stand and a button that allows you to keep it burning like a torch. I turn it on and quickly flame sterilize the tip of the scalpal.
The scalpal is used to quickly open up the stem. The scalpal is used in an area no where near the stem sample. This is simply to open up the stem for quick use. The scalpal was opened directly from the foil it was wrapped in from the pressure cooker.
8) Once you flame sterilize the exacto you can cut the tissue.
8)You can then use your finger on the outside of the stem to pull the mushroom apart in half. Then flame sterilize the exacto to cut the stem tissue. I do this because im only touching the scalpal on the outside of the stem, then i simply open up the rest touching a part of the stem far away from any work im doing on the outside to completely tear the mushroom in half.
9) Use your exacto to cut the inner stem tissue. Quickly lift the blade assembly up and use the inside edge of the jar to push the stem tissue off the exacto into your jar. This entire process took a few seconds so the air exposure was very minimal. Close the lid.
10) Place the black blender base over the blade assembly. You can wipe it down with alcohol or bleach wipes prior to placing it on your assembly. it isn't necessary to pressure cook this. ive taken hundreds of clones this way with no issue.
11) Blend the jar on your oster blender. It will look like this. Blend it until the mushroom particles are very fine. Two quick 3 second bursts usually does it for me.
12) Here is the part I can't show you by myself. Basically quickly lift up your syringe out of the syringe cover, lift up the corner of your metal blade assembly, then suck the liquid up into the syringe. if you have an air gap, flip the syringe upside down and squirt out the air gap so you only see liquid in the entire syringe. Do this very quickly and as soon as you are done sucking in the Liquid Inoculant, cover the top of the blade assembly on the water/stem tissue jar. Also quickly place the syringe back in the cover making sure not to touch the syringe tip on anything. Do this as many times as you can fill syringes. This is for storage later or inoculating whatever you want. You can also use this for Liquid cultures, bulk substrate inoculation, grain inoculation, pf jar inoculation. The blended up syringe tissue will look like this.
You can also pour this blended liquid directly into pre sterilized grain jars. a tablespoon is enough for 1 quart jar. or you can inoculate sterilized substrate in quart jars and fruit directly out of those. i inject 1 10cc syringe into 1 pf jar. it colonizes in 3-4 days, i then slurry to 40-100 quarts which colonize in 3-4 more days. then spawn 10-20 tubs and can be fruiting them in another week. so its very fast.
http://www.shroomery.org/forums/showflat.php/Number/10648149/fpart/1/vc/1
13) Once your done, use a gallon plastic bag and place all the syringes in it. Store in the fridge for future use. Easy as cake.
Here are some leftovers so you can see the blades im using and the stem/clone fruit.
this is enough for clones for an entire year for me. i store them in the fridge. also distilled water is used for the syringes. you can store them directly in the syringe if that is all you have. i simply just use the metal blade assembly as my temporary lid when im cloning . i lift it up and suck up with the syringe. then place the lid sitting back on top of the jar with solution. not tight just sitting on there.
STEM TISSUE GROWING IN PF JARS
ok so im going to illustrate a stem clone taken from Ape strain. it was done using 9er tek and the illustration that was posted in this thread. I did 3 syringes. One syringe was done in 2 pf jars. The other two syringes were done in 4 pf jars each. You can see below how the stem tissue jars that were inoculated with 1 syringe per 2 pf jars is going to explode tomorrow.
Pf jar lids and recipe for pf mixbefore you all flame me for having foil on my lids. its not stopping ge and its not causing water buildup in the verm barrier. i make my mix drier than the normal pf mix so the cakes crumble into small pieces for P2G 123(PF to grain transfer) or Slurry. the lids are illustrated below.
Quote:
my recipe for 24 pf jars
12 cups verm
6 cups brf
5-5.5 cups water. i use a half cup to a full cup less so the pf jars are a little drier. i intentionally do this for liquid culture injections.regular 24 pf jar recipe
12 cups verm
6 cups brf
6 cups waterthis post shows some close ups of my mix. and a description of how i add that last bit of water to the mix.
http://www.shroomery.org/forums/showflat.php/Number/20300997#20300997
here is another post i made with some better close ups of the wet mix after i blend it with a hand mixer.
http://www.shroomery.org/forums/showflat.php/Number/20428297#20428297
ill then use a syringe full of lc and inject one whole or a half syringe into 1 pf jar. this grows very fast with liquid cultures in a few days and then i can g2g that or blend it up and make a ton of jars with a slurry. you may even find that if your going to add a whole syringe to the pf jar, you may want to add 1 cup less than the 6 cups total water for the 24 pf jars. just divide the formula by 24 to get the individual jar ratio.
but if you intend to try the slurry. id inject your lc with a sterile syringe into a pf jar. i usually use a sterile syringe and suck up the liquid culture into it, then inject into a pf jar. im always hesitant adding more moisture to grain jars. but lc's are quick and usually colonize the jars rather quickly. but pf jars are a safe bet. and also, if your only injecting 2 pf jars. your not contaminating 10 quarts if your using a dirty lc. so pf jars for me are also a way of testing if something is clean or not.
Sequence of stem clone growth in pf jars and grain masters. the jar on the far left in the pf jar pictures was injected with 1-10 cc syringe in the jar. it colonized in 4 days. days here is another sequence still growing right now. this was stem clone Liquid Inoculant injected into grain quarts. I did shake it up and I shouldn't have so that delayed the growth quite a bit. I shook it at day 8. this is day 4 thru 10(before Day 1: Day 2: 1 Syringe Per 2 Pf jars 1 Syringe Per 4 pf jars Day 4: 1 syringe per 2 pf jars 1 syringe per 4 pf jars view of all jars Day 5: The bottom shot showing how i colonize the tall 1/2 pint jars from bottom up. i hear all this talk of the tall half pints sucking. well this isn't true you simply squirt your LI or spore solution into the bottom of the jar along the edge of the glass. this colonizes that bottom part first. Day 9: almost done Day 10: 100% colonized. ape is slow. usually other strains will colonize in half the time. if you use 10cc's per 1 pf jar you can get a fully colonized pf jar clone in 3-4 days. you then slurry this into 40-100 quart jars and have those colonized in 3-4 days. you can see how you can get unlimited spawn in a matter of one week. this is almost open air, except it is in the shmuvbox. otherwise, there is no small hole in the lid. its just ghetto. you lift the lid, then suck up water in the syringe. out of 100 syringes...0 contams so its working well. even though a bit ghetto. then you can fill up as many syringes as you want. it may be anal but i always place black electrical tape around my syringes where the lid meets the base. its just an ocd thing i guess. x) Keeping Cultures: Here is a summation of what I do to keep cultures. And it takes up like a small lock box in your fridge that you can keep locked at all times. these will store for 2 years at the least if its clean. i always do a 1 syringe test run in two 1/2pint PF jars and let them grow in 2 days. Speed This is where the speed comes into play. Lets look at it this way, if you have your master sryinges stored in the fridge. All you have to do is inject one syringe into 2 1/2 pint jars. This will grow in a few days with a clone . You then combine this with Tv Casualty's slurry in a hurry tek and you have a clone in a week on grain ready for spawning. Add Tv Casualty's Slurry Tek and.....well there ain't no and. its just fast! How to Colonize Quart jars in 3 Days bernzomatic torch home depot 25$. best 25$ you will EVER spend! guarenteed..best alcohol lamp you can get. had it for years. what this does is preserves your clone . you go back to your original syringes everytime. but if your doing slurry in a hurry to grain. well, then you can use one colonized quart to knock up 12 more quarts for a 2nd generation grain to grain. id stick with 2 generations tops. i had trouble with 3 on a clone with Slurry Tek . this way, if you make lets say 10-10cc syringes. you get enough clone material for a year or so. i can drag one syringe out for months. its quite fast! the beauty of it, is that it takes up no space for storage in the back of your fridge. also, its very quick and simple. you can make one half pint of mycelia turn into 120 jars in a few days using tvs Slurry Tek along with this. fastest way to clone ! highly suggested! original shmuvbox thread from 2003. http://www.thenook.org/forum/ind SOME SAMPLE GROWS USING 9ER TEK CLONING METHOD
anyway, ive been using these methods for more than 5 years with great success. so it does work. if you do it right. all those fancy and complicated supplies are not necessary for the growth of cubensis.
