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Invisibleeatyualive
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Shmuvbox: 9er Tek Cloning and G2g transfers * 9
    #11353633 - 10/30/09 08:05 PM (12 years, 3 months ago)

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.


EZ Shmuvbox Setup: Setting Up A Shmuvbox



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.



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 mix

before 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 water

this 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/index.php?showtopic=2383
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.



SOME SAMPLE GROWS USING 9ER TEK CLONING METHOD




Edited by eatyualive (03/15/16 06:46 PM)


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Offlinecyb3rtr0n
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Re: Shmuvbox [Re: eatyualive]
    #11353730 - 10/30/09 08:22 PM (12 years, 3 months ago)

That's pretty shmuv...                :rimshot:


I take it that you have no problems with the bag deflating and collapsing onto things once you cut the arm holes out?


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Invisibleeatyualive
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Re: Shmuvbox [Re: cyb3rtr0n]
    #11353747 - 10/30/09 08:26 PM (12 years, 3 months ago)

no. the positive air pressure from the hepa keeps it inflated. if you turned off the hepa it would fall. and its taped to the table to keep it in place.


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Re: Shmuvbox [Re: eatyualive]
    #11353770 - 10/30/09 08:33 PM (12 years, 3 months ago)

Makes sense....  So, the turbulent airflow causes no problems for you in the past with petri's and whatnot?  I'm picturing spores whirling around in there if you were working with a print.


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Re: Shmuvbox [Re: cyb3rtr0n]
    #11353847 - 10/30/09 08:49 PM (12 years, 3 months ago)

wow thats pretty sweet gonna have to get me one of those...


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Re: Shmuvbox [Re: cloudsaregathering]
    #11354017 - 10/30/09 09:18 PM (12 years, 3 months ago)



we like it!:super:





good stuff.:sun::plur:


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Re: Shmuvbox [Re: 13shrooms]
    #11354304 - 10/30/09 10:04 PM (12 years, 3 months ago)

also another pointer. keep it on the lowest setting of the airflow. especially if your working with prints.

nice idea with the arm wraps.


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Edited by eatyualive (10/30/09 10:05 PM)


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Invisibleeatyualive
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Re: Shmuvbox [Re: eatyualive]
    #11368167 - 11/02/09 12:31 PM (12 years, 2 months ago)

update


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Edited by eatyualive (11/02/09 12:43 PM)


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Re: Shmuvbox [Re: eatyualive]
    #11514408 - 11/23/09 10:22 PM (12 years, 2 months ago)

i bought the same model.
gonna see how it works in the next week.

anyone else use this?


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Re: Shmuvbox [Re: galadar]
    #11713042 - 12/24/09 06:21 PM (12 years, 1 month ago)

I've been having mixed results myself. So, I'll elaborate... and I do hope that this thread can sort of re-open, as I've read over it like at least a dozen times in the past month. No sense in starting another thread about the exact same thing.

The very first thing I ever used this for was the 9er tek. I attempted to clone a texas adult fruit body. There was a brown stripe running the length of the stipe (I'm a poet and didn't know it :tongue:). I didn't find out that was a bad idea until my brother told me, while reading the 9er tek on here, to me, elbow-deep in the shmuvbox, cloning a suspect fruit.

Great.

Obviously, the cakes I inoculated contamed, with that super bright pink stuff, in case you're curious what it was. However, by the time the pink spot(s) showed up, most of the visible area of the inoculation points were brilliantly white. So, I decided to give it another go with a B+ stipe. This time, no contaminations. ALL of the cakes I inoculated grew well. They have a few small spots of uncolonized area, and should be ready to be made into a slurry in the next couple of days. No suspicious spots whatsoever.

...about a week or so after I made up the first B+ clone, I made another run of 3 fruit body clones, in the 9er tek fashion, once again in the shmuvbox, same sterile procedure, etc. From those 3, one clone contamed almost immediately (I made up 7 WBS jars from these 3 B+ clones, two clones inoculated 2 jars of WBS each, and one clone inoculated 3 jars of WBS). So, I threw those jars out. Then, a few days later, I find that one of the jars, from the clone that inoculated 3 jars, has one whole face of the jar visibly contamed with a dark blueish mold like I've never seen before! The other 2 jars from that clone, however, look and smell great.

There's so many damned variables in those above examples, though. Not to mention the first few times I used it, I wasn't taping the bottom of the bag to the air purifier, or worrying about making the slits large enough, to maintain continuous airflow, out of the eye and arm holes. I've also done a bit of agar work in the shmuvbox now, and that just raises further questions...

I've poured plates in the shmuvbox twice. Once, I stacked all the plates up, let them solidify in the shmuvbox, then placed them into ziplock bags with a spritz of alcohol. The other time, I poured the plates, and placed them into the ziplock bags with alcohol, then stacked them up in the shmuvbox, and let them solidify. Aside from a slight condensation issue (even solidifying in a stack of 20 in the shmuvbox, more than half of them still have condensation on them), not one plate contaminated! I even cleand two old plastic plates with bleach in the shmuvbox, and poured them with success! Now, all of my plates are over 5 days old, and the only ones with contamination are the ones that I've done work with. Well, about a third of the plates I worked with contamintaed. And that to me is the weirdest part. WHY some of my plates have contamed could have a bunch of different reasons, varying from me not getting enough of my scalpel blade red hot (seriously. A couple of my plates contaminated right along all the lines where I cut through the receiving dish to cool it off! How on earth!?), to me not using the shmuvbox in the center of my room, to the wall-to-wall carpeting where I live, etc.

...one last thing before I wrap up this post (sorry it's so long. Hope it's helpful to some:), without going over every last detail of my sterile procedure, and what all I've done in the shmuvbox that's contamintated, etc., I'd like to post some things that I think may explain my varying luck:

1. It just sort of works out that the only desk space I have, is also just adjacent a drafty window, and up against the same wall. I'm thinking that from now on, I'm going to set up a large plastic storage tote in the center of my office/mycology lab/'clean' room, and set up the shmuvbox on that, sanitizing the heck out of it with alcohol and Lysol, etc.

2. My office, where I do my 'sterile' work, is also where I need to fruit things. My apartment sucks, and the only other possible places I could set up shop, would be in either my bedroom (we have 4 cats), or the bathroom. It seems to me that my office is probably the best space I have available.

Well, I hope someone responds at some point, and that this post has been helpful in increasing knowledge of the shmuvbox.


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Re: Shmuvbox [Re: faceyneck]
    #11713051 - 12/24/09 06:22 PM (12 years, 1 month ago)

PM 13 shrooms he can help with any shmuvbox problems


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Re: Shmuvbox [Re: feelfunny]
    #11713064 - 12/24/09 06:24 PM (12 years, 1 month ago)

Thanks, feelfunny! That was speedy!


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Re: Shmuvbox [Re: faceyneck]
    #11718534 - 12/25/09 11:16 PM (12 years, 1 month ago)

Quote:

faceyneck said:
I've been having mixed results myself. So, I'll elaborate... and I do hope that this thread can sort of re-open, as I've read over it like at least a dozen times in the past month. No sense in starting another thread about the exact same thing.

The very first thing I ever used this for was the 9er tek. I attempted to clone a texas adult fruit body. There was a brown stripe running the length of the stipe (I'm a poet and didn't know it :tongue:). I didn't find out that was a bad idea until my brother told me, while reading the 9er tek on here, to me, elbow-deep in the shmuvbox, cloning a suspect fruit.

Great.

Obviously, the cakes I inoculated contamed, with that super bright pink stuff, in case you're curious what it was. However, by the time the pink spot(s) showed up, most of the visible area of the inoculation points were brilliantly white. So, I decided to give it another go with a B+ stipe. This time, no contaminations. ALL of the cakes I inoculated grew well. They have a few small spots of uncolonized area, and should be ready to be made into a slurry in the next couple of days. No suspicious spots whatsoever.

...about a week or so after I made up the first B+ clone, I made another run of 3 fruit body clones, in the 9er tek fashion, once again in the shmuvbox, same sterile procedure, etc. From those 3, one clone contamed almost immediately (I made up 7 WBS jars from these 3 B+ clones, two clones inoculated 2 jars of WBS each, and one clone inoculated 3 jars of WBS). So, I threw those jars out. Then, a few days later, I find that one of the jars, from the clone that inoculated 3 jars, has one whole face of the jar visibly contamed with a dark blueish mold like I've never seen before! The other 2 jars from that clone, however, look and smell great.

There's so many damned variables in those above examples, though. Not to mention the first few times I used it, I wasn't taping the bottom of the bag to the air purifier, or worrying about making the slits large enough, to maintain continuous airflow, out of the eye and arm holes. I've also done a bit of agar work in the shmuvbox now, and that just raises further questions...

I've poured plates in the shmuvbox twice. Once, I stacked all the plates up, let them solidify in the shmuvbox, then placed them into ziplock bags with a spritz of alcohol. The other time, I poured the plates, and placed them into the ziplock bags with alcohol, then stacked them up in the shmuvbox, and let them solidify. Aside from a slight condensation issue (even solidifying in a stack of 20 in the shmuvbox, more than half of them still have condensation on them), not one plate contaminated! I even cleand two old plastic plates with bleach in the shmuvbox, and poured them with success! Now, all of my plates are over 5 days old, and the only ones with contamination are the ones that I've done work with. Well, about a third of the plates I worked with contamintaed. And that to me is the weirdest part. WHY some of my plates have contamed could have a bunch of different reasons, varying from me not getting enough of my scalpel blade red hot (seriously. A couple of my plates contaminated right along all the lines where I cut through the receiving dish to cool it off! How on earth!?), to me not using the shmuvbox in the center of my room, to the wall-to-wall carpeting where I live, etc.

...one last thing before I wrap up this post (sorry it's so long. Hope it's helpful to some:), without going over every last detail of my sterile procedure, and what all I've done in the shmuvbox that's contamintated, etc., I'd like to post some things that I think may explain my varying luck:

1. It just sort of works out that the only desk space I have, is also just adjacent a drafty window, and up against the same wall. I'm thinking that from now on, I'm going to set up a large plastic storage tote in the center of my office/mycology lab/'clean' room, and set up the shmuvbox on that, sanitizing the heck out of it with alcohol and Lysol, etc.

2. My office, where I do my 'sterile' work, is also where I need to fruit things. My apartment sucks, and the only other possible places I could set up shop, would be in either my bedroom (we have 4 cats), or the bathroom. It seems to me that my office is probably the best space I have available.

Well, I hope someone responds at some point, and that this post has been helpful in increasing knowledge of the shmuvbox.




turn off your ac when you do clean work. i turn it off 30 minutes prior to any clean work. ive done 5 years worth of 9er tek clones in a shmuvbox with maybe 1 possible contam. it was eq strain. and it actually just didn't germinate. other than that. if you do your work quick with good technique. and don't touch any stem tissue or anything on the outside of the jar with your syringe. it works like a charm.

you also want to pre sterilize everything in your pressure cooker like utensils. anything you put in that bag you want to wipe with alcohol. and did you wipe the filter prior to doing any work? clean it, wipe it down with alc. i then spray some lysol in the bag. let it air out for a few minutes prior to lighting anything or doing any work in it. anytime you take your hands out of that bag. make sure you wipe your arms, your hands, or put gloves on b4 placing them back in the bag. make your work clean, fast and you should have no issues. sounds like you need to straighten some things out on your end first. designate an area. a table works. buy yourself a plastic table cloth. lay that same cloth over your clean area. wipe it down with bleach wipes, rubbing alc. turn off your ac. wipe down all utensils. be super fucking anal about it. like one germ gets in there and your fucked. think of it that way. i just do it on a folding table from office depot. once your done with the clean work. fold it up and put it behind the couch or in the closet. fold up your table cloth. and only use it for clean work only. only use the filter for clean work only. put the filter up each and every time you use it. put it in a plastic gallon bag. i go through approximately 1 filter every 3 years with this setup. and i only use this filter for clean work. solely. it does not run but maybe 30 minutes total a week if that. recently i have not done any clean work besides g2g's in the shmuv bc it wasn't necessary. but g2g's work fine.

you need to take the time to get your transfer tek down quick. anytime you open a jar you want to close it asap.

anytime you transfer stem tissue you want to make sure it doesn't touch anything. sterilize your utensils with a bernzomatic torch prior to cutting tissue. use an alcohol wipe to rub on the utensil after you have sterilized it to allow it to cool prior to cutting your stem tissue. make sure when you open up the jar, nothing touches the lid or outside of the jar. keep it all inside the jar with the clean water and you should be fine. you can set your fruit in the shmuv for a long time. your main concern is once you open up the stem, to make the transfer and airtime minimal.


did you turn your hepa to the lowest setting when you did your work? thats a key point. very slow airflow. not heavy works great.

ive poured plates with no problems. but go the extra mile to make sure your not contaminating things by dirty hands.


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Re: Shmuvbox [Re: eatyualive]
    #11719788 - 12/26/09 04:15 AM (12 years, 1 month ago)

Sweet! Just the person I wanted to talk to! I REALLY want this to be my one and only setup for sterile procedure. I just love working with it so much. I've had outrageous amounts of contams with it recently of course, but fortunately, I have many areas that I can improve on.

Quote:

turn off your ac when you do clean work. i turn it off 30 minutes prior to any clean work. ive done 5 years worth of 9er tek clones in a shmuvbox with maybe 1 possible contam. it was eq strain. and it actually just didn't germinate. other than that. if you do your work quick with good technique. and don't touch any stem tissue or anything on the outside of the jar with your syringe. it works like a charm.

you also want to pre sterilize everything in your pressure cooker like utensils. anything you put in that bag you want to wipe with alcohol. and did you wipe the filter prior to doing any work?




I haven't paid much attention to my ac. I don't think it's turned on while doing sterile work, however, it may have. I'll be sure to turn it off now.

...as for cleaning the HEPA filter prior to use: I wipe every outside surface of it down with alcohol, and then spray a bit of Lysol onto the scrubber filter thingy. You know, the black stuff that is inside the plastic grate, covering up the filter. Then, I spray shitloads of alcohol into the shmuvbox bag, run it on the lowest setting, with the ionizer on every time, so far, and then spray shitloads of oust into the bag, and then cloud the room with oust, and let it clean the air for about 10-15 minutes.

Well, I'm going to be using my shmuvbox, in the center of the room that I've been doing my work in this time, and I'll also throw down a plastic table cloth, as well. I'll also start storing the HEPA filter in a bag, with a spritz or two of alcohol.

Quote:

did you turn your hepa to the lowest setting when you did your work? thats a key point. very slow airflow. not heavy works great.

ive poured plates with no problems. but go the extra mile to make sure your not contaminating things by dirty hands.




Yeah, I've always used the lowest setting. However, I now have a few plates, that have some contams in them. Those plates, I should transfer in a glovebox, right? Or, maybe prop up a trash bag with the help of the the shmuvbox, and then turn the flow of air off?

Also, do you concern yourself with where you're working within the shmuvbox? Like, should I keep the items I'm working with near where the filtered air comes out of? Or, should I work near the hand holes? Away from the hand holes? Or, what?

Thanks again, EatYu! I've been following your posts quite prolifically lately. I have a bunch of cakes that are now fully colonized, from my first successful 9er clone, that I'll be making a slurry with. I'll be inoculating jars of WBS, as well as some of my signature substrate filter patch bags, with the slurry. I have a feeling it's going to be magical! And, growing in this way, is what I want to ultimately do exclusively. So, it's CRITICAL I figure out how to stop screwing up the damned shmuvbox!

Super-Dee-Duper-thanks-ez. However; please don't eat me! Or my shrooms! I DEFINITELY won't be able to perfect the shmuvbox technique then!

Facey

P.S.: 13Shrooms recommended that I keep the inside of the bag dry while working in it. Also, as I mentioned before, the only thing that I've had 100% success with, is pouring plates! And, of course, the inside of the bag, was completely dry, while pouring plates, every time. Other teks; not so much.


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Re: Shmuvbox [Re: faceyneck]
    #11720683 - 12/26/09 12:00 PM (12 years, 1 month ago)

this is how i do it.

tape the bag around the unit, neatness doesnt matter just make sure its airtight, itll just inflate into a ball thing anyways. dont cut yet.

seal windows cracks w.e
bomb room with clean linen scented oust in a backwards motion exiting the room :smile: for 20 seconds just enough for a little cloud. [1:9 bleach:water in a spray works great too]

give it a minute or two while i throw on surgical mask, gloves, cover hair etc.

go in bomb the air a little more, idc you gotta be anal like eat said.

spray or wipe hepa unit all around, open it up, spray a puff or two inside,
take hepa filter and prefilter out of ziplock bag that i use to store them in. and spray them down and wipe then assemble the unit.

turn on unit to low setting, with the ionizer on.
bag will inflate, now take a blade or exacto or scalpel[that obviously isnt gonna be used for agar or cloning :wink:] and slice two 5-6" horizontal holes for your arms.
If youre using clear bags, no need for an eye hole.

get inside, spray down the insides especially the bottom part of the bag.
load jars in, load tools wrapped in foil from pc'ing them in the shmuvbox.

spray a puff inside, and in the air. get your hands wiped all over with alcohol and begin work!

placement in the box doesnt really matter. just make sure you keep that filter use to a minimum!

every time youre done, get rid of the bag, take out the prefilter and the hepa and store in a plastic bag with a puff or two of oust.  and without the filter the hepa is basically a fan that you can keep running for fruiting.


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Re: Shmuvbox [Re: faceyneck]
    #11722148 - 12/26/09 06:47 PM (12 years, 1 month ago)

Quote:

faceyneck said:
Sweet! Just the person I wanted to talk to! I REALLY want this to be my one and only setup for sterile procedure. I just love working with it so much. I've had outrageous amounts of contams with it recently of course, but fortunately, I have many areas that I can improve on.

Quote:

turn off your ac when you do clean work. i turn it off 30 minutes prior to any clean work. ive done 5 years worth of 9er tek clones in a shmuvbox with maybe 1 possible contam. it was eq strain. and it actually just didn't germinate. other than that. if you do your work quick with good technique. and don't touch any stem tissue or anything on the outside of the jar with your syringe. it works like a charm.

you also want to pre sterilize everything in your pressure cooker like utensils. anything you put in that bag you want to wipe with alcohol. and did you wipe the filter prior to doing any work?




I haven't paid much attention to my ac. I don't think it's turned on while doing sterile work, however, it may have. I'll be sure to turn it off now.

...as for cleaning the HEPA filter prior to use: I wipe every outside surface of it down with alcohol, and then spray a bit of Lysol onto the scrubber filter thingy. You know, the black stuff that is inside the plastic grate, covering up the filter. Then, I spray shitloads of alcohol into the shmuvbox bag, run it on the lowest setting, with the ionizer on every time, so far, and then spray shitloads of oust into the bag, and then cloud the room with oust, and let it clean the air for about 10-15 minutes.

Well, I'm going to be using my shmuvbox, in the center of the room that I've been doing my work in this time, and I'll also throw down a plastic table cloth, as well. I'll also start storing the HEPA filter in a bag, with a spritz or two of alcohol.

Quote:

did you turn your hepa to the lowest setting when you did your work? thats a key point. very slow airflow. not heavy works great.

ive poured plates with no problems. but go the extra mile to make sure your not contaminating things by dirty hands.




Yeah, I've always used the lowest setting. However, I now have a few plates, that have some contams in them. Those plates, I should transfer in a glovebox, right? Or, maybe prop up a trash bag with the help of the the shmuvbox, and then turn the flow of air off?

Also, do you concern yourself with where you're working within the shmuvbox? Like, should I keep the items I'm working with near where the filtered air comes out of? Or, should I work near the hand holes? Away from the hand holes? Or, what?

Thanks again, EatYu! I've been following your posts quite prolifically lately. I have a bunch of cakes that are now fully colonized, from my first successful 9er clone, that I'll be making a slurry with. I'll be inoculating jars of WBS, as well as some of my signature substrate filter patch bags, with the slurry. I have a feeling it's going to be magical! And, growing in this way, is what I want to ultimately do exclusively. So, it's CRITICAL I figure out how to stop screwing up the damned shmuvbox!

Super-Dee-Duper-thanks-ez. However; please don't eat me! Or my shrooms! I DEFINITELY won't be able to perfect the shmuvbox technique then!

Facey

P.S.: 13Shrooms recommended that I keep the inside of the bag dry while working in it. Also, as I mentioned before, the only thing that I've had 100% success with, is pouring plates! And, of course, the inside of the bag, was completely dry, while pouring plates, every time. Other teks; not so much.




actually take the filter out of where it sits. wipe that whole area with rubbing alc as well. i also spray a little lysol inside the area where the mechanism is. not a heavy dose of it. but enough to do the trick. i also wipe down the outside of the hepa filter itself. the plastic part. i don't wipe anything on that scrubber your talking about. but i do wipe down all plastic parts of the filter itself. as well as the housing that holds the filter and the black scrubber thing in. this area is wiped inside and out. then i wipe the outside of the whole unit in the entire area that will be going inside the bag with rubbing alc.

also remember. any jars your adding or anything into that bag. you need to wipe down with an alc wipe or bleach wipe prior to placing it in the bag. but it sounds like your doing things on the right track. try to minimize your use. ill often do max 2 clones at a time via 9er tek. i try to keep it minimal. it can be tight to work in so you want a little breathing room so you don't knock things over. just do one clone at a time until you get the proper tek down. then once you have it down it will be a piece of pie. for me this method actually has worked since the first time i tried it. but ive also had work with flow hoods and things like this prior to using one.

yeah i just mean, do it in an area that is draft free. not right under a house or apartment cooling vent. turn the ac off. then do what you said. sounds like your ousting away. thats a good thing.

i usually do the work right in front of the hand holes. any time i open something sterile i minimize airtime. yes, you don't want to open any contams inside it. that will push them everywhere.

make it easy. just wipe down everything that will touch the inside of that bag. that includes the area where the filter takes the air in. the part of the hepa unit that holds the filter in, the inside mechanism that sucks the air in and the part of the hepa unit that will have the bag taped over it. wipe it all down and you should be good.


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Edited by eatyualive (12/26/09 06:59 PM)


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Re: Shmuvbox [Re: eatyualive]
    #11722165 - 12/26/09 06:52 PM (12 years, 1 month ago)

several things I could do to improve my sterile schmuv tek posted here.  good info :thanx:




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Re: Shmuvbox [Re: 13shrooms]
    #11722181 - 12/26/09 06:55 PM (12 years, 1 month ago)

also lets say you do spread a huge contam into your filter. well. just get a brand new hepa filter. not the entire unit, just the hepa itself. yeah it might cost you but better to be safe than sorry. ill usually buy the hepa unit. and 2 extra filters. that i still have the filters from several years ago that were purchased with this unit. still on filter number 2. have one left. had one batch of contams this year. and its over with now. but not bad for a whole year. it was a huge hit but managed through it. had to really wipe the hell out of the shmuv. had a cake was about to go to g2g into grain jars. forked it and green went everywhere. tossed out all the grain jars even though they were all closed. also wiped the shmuv inside and out. 3 times over. wiped the whole room. didn't do any clean  work for like 2 months just in case lmao...


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Edited by eatyualive (12/26/09 06:56 PM)


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Re: Shmuvbox [Re: eatyualive]
    #11724213 - 12/27/09 03:45 AM (12 years, 1 month ago)

Shyeah, I know what you mean! Can't be too careful!

...and that's of course what's frustrating me. I'm pretty anal about being clean and careful and stuff. I'll definitely start using the shmuvbox for smaller loads of sterile work at a time, now. I'm sure that'll help, at least at first.

What's the longest you would say you've worked in the shmuvbox in one sitting? What do you think is the longest length of time one could still safetly get everything done in?

I'm definitely going to buy a new filter. I have a feeling my present filter isn't exactly great. All of the units, at Wal-Mart, when I bought mine, were super dusty. Been sitting there a while. Also, I've had CRAZY HUGE trich outbreaks (had a faht jar with some trich hidden in it. Know idea how it happened. The culture looked PERFECT on grain, but once I finally transferred a grain to agar to test it out; NOTHING but trich!), not on anything out in the open air, but I did end up growing 8, 5 pound, large filter patch, substrate bags, of the MEAN GREEN!! :suckit: I hate you trich!! Everything necessarily takes place in the same room at the moment. Bummer.

So, the very few trays I have left, are on their second and third flushes. Do you think, if I tried to do a 9er clone, and am successful, with a 2nd flush fruit, that I'd LIKELY be able to grow a full canopy with it? Or, would there be a likelyhood, that I'll grow a bunch of trays that only ever fruit with like 2nd+ flush density?

Thanks again, Eats.


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Re: Shmuvbox [Re: faceyneck]
    #11728787 - 12/27/09 11:50 PM (12 years, 1 month ago)

sometimes ill be in it for an hour. if im worried. and i do a 2nd sitting. i start fresh. like new bag, clean the filter the whole 9 yards. plan this ahead with utensils. i usually have two scalpels pced prior so i can do the double work. this also includes 2 9er tek blender jars as well.

remember working with clones off fresher material is always best. try to clone off 1st flush off 1st generation spawn if possible it will last longer. after multiple transfers and multiple clones you can see the degradation. the mycelia won't catch on grain. the growth is slow. substrate runs look healthy but a bit slow. then end up contaminating  a few days into fruiting conditions. i try not to clone a clone more than one time.

ive also cloned 3rd flush fruits with mixed results. ive also done it in 2nd flushes off year old clone material and had good success. but the older it is. the weaker it will be in most cases.

tasmanian strain is one of those strains that last a really long time in the fridge. got a 2 year old clone fruiting like crazy still.


some other trials if your interested.

http://forums.mycotopia.net/fungi-growing-edible-medicinal-magic-mushrooms/61337-shumvbox-trials-refining-my-shumvbox-tec-log.html



--------------------
EAT GETS SHIT DONE


:flame::chief:JOIN THE POW WOW:chief::flame:


Edited by eatyualive (12/29/09 12:22 AM)


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