Mephisto's Master Slant Tek

Discussion in 'MYCO-TEKS' started by Mephistophelian, Apr 2, 2012.

  1. Mephistophelian

    Mephistophelian Six Legged Pilot Mushroom Doctor

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    Its easy...doesn't take special equipment or nonsense and is very straightforward. It also works with any type of slant you may take interest in be it PP Screw Cap, Simport CulTubes, Pyrex Screw Cap, or Vac-Tainers. This works with them all...especially if you don't have a sterilizer setup or a test-tube rack.

    Tube Measurements:

    • Vac-Tainers - with a red top are commonly 10ml and 16mm x 100mm
    • CulTubes - depends on your model, I used the largest being 17mm x 100mm (They are also PP)
    • Absolute Smallest Recommended Dimensions is 16mm x 100mm, no less.

    So lets begin.

    Materials Needed:
    • Wood Sticks - Popsicle, Match Stick, whatever type you find that is flat and suitable for the job
    • Slants/Tubes w/ Caps- Vac-Tainer, CulTubes, Pyrex or PP
    • Agar Mix - I used Lab Standard MEA
    • Measuring Container - Mix agar
    • Syringe - Whatever size & gauge you have laying around.
    • Pint Jars - Quart jars would work as well, although I wouldn't want to personally do it with Quart jars
    • Cooking Pot - Any size is suffice Optional
    • Scale - For measuring agar mix (unless you have premix)

    [​IMG]

    [​IMG]

    Step 1: Preparation

    First thing, measure your sticks to see if they fit sufficiently. I had to break all my sticks in half lengthwise then in half width wise to fit right. Do this before anything else. Pre-soak your woodsticks. I boiled some water in a pot and dropped them in. Took it off the stove, covered with a lid and let soak. You can move onto step two while you wait.

    [​IMG]


    Step 2: Preparation Continued

    Fill your slant with water WITH a stick in it, to approximately 7ml. No more then that. As low as 5ml but no more then 7ml WITH the stick in it. Without the stick, it will vary by .5ml. You need this to be no more then 7ml because when you go to lay your slant down, you will lose surface area for your mycelium to grow. That's no good. You'll see when you pour your water in and slant it so the water doesn't go past the edge (remember that the lid will take up a small space inside the tube and you don't want agar building a wall up against it inside).

    [​IMG]

    Now, you want to go into your Lab/Room and inside your GB or in front of your flow hood, have your spot where your slants will sit while they cool before hand. It sucks to do it while your unloading the jars and getting it ready last minute. Not wise. We see here that my little racks sitting on 4 popsicle sticks was suffice to make the perfect slant.

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    Now that we have that setup, lets go check on our sticks sitting in the hot water.

    Step 3: Sticks 'An Tubes

    [​IMG]

    Looks like the sticks are ready and saturated. In a pinch, you can soak these overnight instead of hot soaking, but hey...sometimes you forget or get an urge to do something right-this-second and well hot soak works well. Now what you do is get your tubes ready and lay them out and put your sticks in them. Once you've got your sticks laid out, and sticks in them. We can go to the next step. See how easy this is? You could be drinking a beer while doing this. But make sure your still on your first by this point otherwise your in for a world of trouble later. ;)


    Step 4: Agar Fun

    Now lets get some agar mixed up. Some prefer to do it in a pot with it being warmed, some do it cold. Personally I do it with warm water poured into the mix but not over a pot. Had some of my agar go jelly in the pot one time, never bothered with it since. I've got enough temperamental things in my life.

    The agar recipe I used was the standard formula of 2% nutrient base 'lab standard' from Fast Freds Cookbook.

    10G Malt Extract
    15G Agar
    1L Water

    For myself, I used 300 ml of the agar for this project (the picture it shows just above 500ml with displacement of the powders the rest of the agar was used elsewhere) which makes the recipe cut in half. What you need to do is decide how many your pouring and do the math. 7ml x (how many slants) = total water needed. Adjust nutrients and agar to suit. Simple no? Here is my agar & ME on the scale.

    [​IMG]

    Now stir it all together.

    [​IMG]

    Time for the syringe. Stir up your agar mix to get all the powders that aren't dissolved and suck up into the syringe and start to fill your slants with maximum 7ml a piece. This is simple enough. If you happen to bugger off and come back a few minutes later, make sure to shake your syringe around to get the powders sloshed about to make an even mix for each slant.

    Again, STIR BEFORE DRAWING SYRINGE

    If you have CulTubes, there are two snaps. First snap is aerobic and lets air through, press harder and its air & water tight. Just leave it loose so air can get in. They'll pop off if you don't. For Vac-Tainers see attached picture. For Pyrex screw tops, just leave it slightly unscrewed to let air through. Don't worry about contams right now, just keep reading the guide, we'll get to how to avoid those.

    [​IMG]

    Get a pint jar to start to lay these in. You'll fit no more then 8 using CulTubes, I think it'll be 8 with Vac-Tainers as well. Grab two lids of whatever type you decided on and lay them down then lay your pint jar down on them so you can slide the tubes in easier.

    [​IMG]

    Repeat this until all tubes are filled up and all your jars are full.


    Step 5: Preparing Jars for PC

    Now I don't know if this is absolutely necessary but, I like to take precautions and would not want my newly poured slants to get all 'splody in my cooker while they're cooking. So I fill my jars up with some water just about to the same level as the agar in the tubes. I try not to go too high since that water may get up into the slants and throw the content off some. Your welcome to follow suit or do your own thing...don't matter to me. I'm just showing you how I get it done.

    [​IMG]

    [​IMG]

    Now that all your jars are filled with a bit of water and all your tubes are capped (loosely). Now its time to put on your grain lids that you use in all your grain jars. Tyvek filters, synthetic filters, whatman custom lids, whatever...you just need some filtered G/E like you would with grain for these. This lets air pass through and around it all without needing a sterilizer to vacuum seal the whole ordeal.

    [​IMG]


    Step 6: PC & Cooling

    Toss them all into the PC now. I do it for 15-20 minutes at 15 PSI which is suffice for agar. I fill my PC to match the height of the water in the jars to help slow down the cooling in the jars and slants. This helps reduce or eliminate any boil over you might find. Once they are all PC'd and the PC cools enough to open it up, take your jars up to the Lab/Room with your Hood or Glove Box.

    Treat these jars like you would with grain jars. No different, keep it all clean and good. Now, open your jars up and when you open up the jars the first thing you do with the slant you pull out is to either snap the lid down tight (CulTube/Vac-Tainer) or screw the lid tight (Pyrex/PP Screw top). This way you stop contams from getting in while they cool. Then lay them down so your wood stick is on the bottom of the slanted agar and let them cool in that angle till your ready to go. They will stay sealed and contam free for a long long time at room temperature. I still have slants that are in VacTainers that I didn't use and are still sitting in my drawer clean and problem free over a year later. On the note of vactainers, when your inoculating them, be sure to keep the vent hole up. Its kind of a pain, but its absolutely necessary (with either glad-wrap or parafilm, both work). But you gotta have the vent up.

    [​IMG]

    [​IMG]

    And there you have it. Slants to use for your precious cultures or for precious isolates of cultures your studying and working with. You don't even need petri dishes to transfer cultures to these (although it is highly recommended). On a side note, I did have a couple of my slants have half the water/mix in them then they did when it started. It happens every time I work with these and its only 3 or 4 (even within a jar of other full well made ones). So if it happens to you, don't be bummed out. One thing for the nay-sayers out there for the method of the agar, as I finished typing up this guide I went to check my tubes, all jelled and hardened nicely as they always have without heat treatement or 'heavy mixing' like some folks say you have to do.

    I hope you enjoyed this and that it will suit you well for many many years. :sun:
     
  2. tripdawg420

    tripdawg420 Active Member

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    :awetongue:nice tek you made it look so easy
     
  3. EvilMushroom666

    EvilMushroom666 Well-Known Member Moderator Mushroom Doctor

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    Thanks for sharing this amazing TEK with our community brother, much apprecaited:thumbup:
     
  4. Weller

    Weller Student of Nature Moderator Cannabis Doctor Cacti Doctor

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    Thanks for sharing. I will definitely use this tek when i make my 1st slants :super:
     
  5. HorizonSpawn

    HorizonSpawn Moderator Moderator Mushroom Doctor

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    This is THE tek I use for making master slants :thumbup1: Nothing can best it, for a SAB user :dance1:
     
  6. Toni

    Toni Moderator Moderator

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    This tek is helpful for most mushroom growers, hats off to your contribution to mycology :hatsoff:
     
  7. Professor PinHead

    Professor PinHead Lost in the Tek.... Administrator Mushroom Doctor Cannabis Doctor Supporter

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    Thanks for posting brother!

    This is one of the greatest methods I have seen for prepping slant hands down! :thumbup:

    Still Air Box or not, lol.
     
    Down311 likes this.
  8. Mephistophelian

    Mephistophelian Six Legged Pilot Mushroom Doctor

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    I would hate to have to do this in a Glovebox, I would want to rip my hair out. But I wanted to make a Tek that was friendly for both sides of the fence...since not everyone can flesh out $300 on a flowhood.
     
  9. Professor PinHead

    Professor PinHead Lost in the Tek.... Administrator Mushroom Doctor Cannabis Doctor Supporter

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    My hood was 1000, :hehehe:

    My first one was about 400 and some change......... :thumbup:

    I have done my fair share of hair pulling to get there though don't get me wrong.
     
  10. Jeff

    Jeff Well-Known Member Moderator Mushroom Doctor Supporter

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    This is the tek that got me started making slants. Great write up and thanks for bring it here.
     
  11. OoBYCoO

    OoBYCoO Super Moderator Moderator Mushroom Doctor

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    Archived! :tony1:
     
  12. mycborg

    mycborg Well-Known Member Supporter

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    Dam nice :super: I definetly gonna do this approach next round!
     
  13. Mephistophelian

    Mephistophelian Six Legged Pilot Mushroom Doctor

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    Thanks for the kind words :thumbup1:
     
  14. sage_vyasa

    sage_vyasa New Member

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    this may be a bit off topic, but is it possible to culture these fungi with a deep stab technique? I have more microbiology background than mycology and know that deep stabs can limit GE somewhat.
     
  15. Professor PinHead

    Professor PinHead Lost in the Tek.... Administrator Mushroom Doctor Cannabis Doctor Supporter

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    You mean like a punch boiopsy?

    You don't want that. In fact even if you did that the mycelium would climb out of the trench and would still end up covering the surface of the agar media.....
     
  16. insectocean

    insectocean Stranger Than Fiction Supporter

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    I just moved today and it felt like my hood weighed 1000, down one flight of stairs across town and up another :wow:

    Mephisto: Does PP hold up to multiple pressure cycles as well as pyrex or glass? Great tek!, I need to make up a run of culture slants.
     
  17. Mephistophelian

    Mephistophelian Six Legged Pilot Mushroom Doctor

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    I haven't seen PP have any issues with multiple pushes on it since it can handle these specs:


    Maximum Temperature: 275°F 135°C
    Minimum Temperature: 32°F 0°C
    Autoclavable: Yes
    Melting Point: 338°F 170°C
    Tensile Strength: 4,500 psi
    Hardness: R95
    UV Resistance: Poor
    Translucent
    Rigid
    Specific Gravity: 0.90
     
  18. Larynx

    Larynx Well-Known Member Mushroom Doctor

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    I found the quality of PP varies between sources. Low grade PP tends to morph after repeat using. The higher grade stuff works like a champ.
     
  19. Mephistophelian

    Mephistophelian Six Legged Pilot Mushroom Doctor

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    Interesting to know Larynx, thankfully I haven't encountered poor PP as of yet. I think in terms of Culture tubes and the like...they would (one would think) be higher quality.
     
  20. FractalXplora

    FractalXplora Master Mycologist

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    Great tek Meph, :naughty:this is also my touchbase tek for when I learned this fine art of slanting, just had a reread of it to refresh my memory,glad I did as I forgot all about putting them in filtered jars with water!

    Frac.
     
    Last edited: Aug 8, 2016