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InvisiblefahtsterM
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Cloning fruits via Josex’s poke * 9
    #28301297 - 04/30/23 04:02 PM (8 months, 25 days ago)

Disclaimer: as a lot of you know, I tend to get wordy in my procedural threads.. this isn’t because I like hearing myself talk, it’s because I try to cover any and all questions that might come up, in the OP.  I’m not always around to answer questions.  I’ll post the abridged version first so you guys can get the gist and you can click here for the extended version where I go into more detail

First off I’d like to give credit where it’s due.  Josex’s poke is the basis for this method with the main difference being that I’m poking inner cap material using a modified 16g needle and then injecting that through a SHIP into a small 4oz grain jar

Abridged Version

1. Zip tie foil over lid of 4oz grain jar.  Position a “lip” of foil in front of SHIP.



2. Modify 16g needle by cutting most of the tip off but keep it sharp.  Cut like so..



3. Put empty syringe with modded needle into a 4oz jar filled with water and cover tightly with foil



4. Pc both the covered grain jar and syringe jar for 2hrs @ 15+psi



5. Once cool, open the pc and immediately pull the syringe plunger to fill the syringe and then tighten your grain jar lid

6. Cut your cap off most of the stem, leaving a couple inches of stem so you can pull the cap in half cleanly

7. Cut a slit in the foil just above the zip tie right in front of the SHIP

8. Flame your needle red hot and set down

9. Pull the stem towards the cap to split it in half exposing inner cap tissue

10. Grab your syringe which should still be hot and squirt a drop or two out to cool the needle.  poke the inner cap tissue a couple times to get a myc plug in the needle barrel

11. Pull tab of foil on your grain jar (or LC jar) to expose the SHIP

12. Inject the needle with myc plug into the SHIP and push the plug in

Done!



Couple of plugs starting to jump off



Some jars colonizing after a/few shake(s)



If you’re doing multiple clones/jars, just reflame the needle.  You can easily do 5+ clones per syringe.

Lots of clones to test!  Haven’t lost one yet



I’ll update this with any clones I grow out using this method

Here’s PE clone I got using this method the first time I tried it last year (inner cap needle biopsy)



Mak x yeti f1 clone (this was pre-needle modification.. I poked through the other side of the cap on this one so not sure what caused the first flush to grow like it did—could also have been lack of water to support the flush because the second was nice)



PE/RW (Charlie Brown) f7 clone



Fp+ f8 clone.  These guys really don’t do well in summer months due to how much extra heat the sub produces because of how dense the myc grows.  For most of this tubs fruiting, the temps were 80F+.  I still got 52g dry so I’m going to redo the grow in the fall because I think this can be a killer culture… love the pheno


Here’s 2nd flush coming in.. I’ll probably end up with ~70g dry which isn’t horrible for a sub that crapped out


EDIT 1.25.24:  redid a grow with this culture and it turned out pretty nice.  I used a peat casing to reduce standing water beads (I think the summer heat coming on matched with the water beads sitting in the surface of the sub for 2+ weeks turned the surface bacterial the first tub).  Dried down to 79g first flush



Here’s a fp+ f9 clone I took off a second flush qt bag colonizing its 4oz grain jar… it’s being dried out for storage to grow in the fall as well.  This clone was put to a grain jar that had been wrapped in syran wrap for a 25 days (as you can see, the grains are still well hydrated) and I used the steam pen to clean up the SHIP.  <— not my preferred method but it works


Since the needle mod, all the clones have recovered/grown beautifully


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InvisiblefahtsterM
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Re: Cloning fruits via Josex’s poke [Re: fahtster] * 1
    #28301312 - 04/30/23 04:21 PM (8 months, 25 days ago)

Extended Detailed Version

Yoyoyo, party peeps! 

So some of you know that I’ve used Oven bags to clone and it’s treated me well since I’ve done it but it’s got it’s limitations.  I can only do a couple clones per bag.. it’s bulky and there’s a lot of moving parts.  Prep is much easier with this newer version as well

Easy cloning was kind of my last hurdle without using agar.

It’s not really a new idea or anything but there’s a couple things that make this extremely easy and reliable that I think it’ll help a decent amount of ppl.

One of the big factors that make this go off well is that I eliminated the need to wipe the SHIP before poking.  More on the in a second.

The other big problem when trying to poke fruit material is that the stipe tissue is usually really stringy.  The reason that’s a problem with this method is because even if you do get a “plug” of myc in the needle barrel, it’ll just get pulled out again because it’s still attached to the rest of the “string”

The fix for this is to use inner cap tissue instead of stipe material.  It’s not stringy at all and is more like tiny orbs of myc stuck to each other and there’s no problem getting a myc plug to stay in the barrel.

Another downside to poking cap material with a normal hypodermic needle is that it’s too long from the sharp tip to where the actual opening to the needle barrel is… more times than not, if you try poking far enough into the cap to actually retrieve inner cap material, you’re probably poking through the outside of the cap on the other side and possibly contaminating the very tip.

The fix for this is to use a small very sharp wire cutters and cut most of the tip off while still keeping it sharp at the end.  You can use a blunt needle to inject through a SHIP but it takes a decent amount of pressure and you risk picking up a plug of silicone (in my case).  Keeping the end still sharp and pointed keeps that from happening.  Cut like so..



Now that you have your modified needle, onto the modified lid… this is really simple.  Basically I zip tie a piece of foil over the lids and SHIP.  The foil cover has a little lip of extra foil positioned in front of the SHIP so that when I’ve got my needle tip with a myc plug, I can easily pull that tab to briefly expose the SHIP and immediately inject the needle into the ship without having to wipe it with anything and possibly contaminate it.  The zip tie is to keep any air out so the SHIP remains clean after the pc cycle until you’re ready to clone



**I wish I could take credit for the foil idea (certain parts I came up with like the pull tab and zip tie but someone mentioned the foil cover idea in passing a while ago and I thought it was a great idea so kinda put it in the back of my mind—if that was you and you remember the interaction, please speak up!)

So that’s pretty much the prep that’s needed to do one clone (minus the grain prep—in my case—or LC if you’re going that route).  The cool thing is that one syringe filled with sterile water can do many clones.. all you do is flame the needle between them… sometimes the plug gets a little stuck and you end up squirting out a bit more than other times but I can easily do 5+ clones per filled syringe and it’s fast.

I guess I should show the lids I use for this… I use a large DIY SHIP but any secure lid with a SHIP will work.  Here’s my lid set up

Let’s get to the procedure, yeah?

Preparing your materials

1. I use 4oz grain jars (wheat is my preferred grain).  Prepare them as you would any grain jar (2hrs @ 15+psi). 


It’s hard to tell but they’re filled 2/3 the way so there’s room for shaking.  Any less grain and they can dry out too fast before the plug has a chance to really take off and you get full colonization (although, it’s not necessary for them to be fully colonized… in my case, these jars are only for myc expansion.  Even if they are only half colonized, that’s plenty of myc to GLC to a couple qts and another 4oz clone extension jar.  Those will colonize much faster because of you’re using LC and not waiting on a tiny myc plug to expand)

The cool thing about these 4oz jars is that I can make a bunch of them in one pc run and afterwards, just wrap them in cling wrap (to slow/stop them from drying out) and just grab one when I need to clone.  I’ve used them 3 weeks later and saw no visible drying out of the grains even on the top

EDIT FALL 2023: this ^^^ was a bad idea… at least for how I make my lids (use to now).. I used tape on my filters and that tapes adhesive will grow mold when the FAE/GE is cut off like that with wrapping in syran wrap.  I didn’t notice it at the time of taking the clones but months later when I tried reviving the dried out clones I made with those jars months later, it was mold city.  So for now, I’ll be making the clone jars and using them immediately.  So, don’t do this vvv if you’re using a filter on your jars that shouldn’t get wet



When I do use them later, I just attach the foil and pc again for 30mins.. this is only to sterilize the SHIP under the foil again.  You could probably even get away with bringing the pc up to pressure (15psi) and then just turning the burner off… you’re not sterilizing anything but the outside of the jar (ie. SHIP and maybe a syringe with water)

2. Attach your foil cover to the lid using zip ties.  Depending on how big they are, you may have to put a couple/few together so it can reach all the way around your lid.  Make the foil a decent amount bigger than the lid size so that on one side of the foil piece, you can fold it enough to make a tab that you can pull with your fingers to rip the foil back and expose the SHIP right before you inject the myc plug.  Make the tab right in front of your SHIP



**obviously, if your using the jar right away, attach the foil before you pc for the 2 hours… don’t forget to loosen the lid a 1/4 turn and tighten when you take it out of the pc

That’s it for the jar prep.  I’ve also used the Steam pen instead of the foil (especially useful if I’m using jars that were presterilized) but I worry that the heated up SHIP will damage the myc plug but it does work… it’s more of what I feel like doing at the time

3. Filling the syringe.  With the modified needle/syringe I fill them by setting the syringe in a 4oz jar of water (plunger all the way in) and cover with foil like so..



I have a 16qt presto and it isn’t tall enough to set a filled syringe in a qt jar of water.  So the first thing I do when I open the pc is hold onto the syringe barrel and pull the plunger up to fill the syringe with the sterile water.



Now that you have your filled sterile syringe and foil covered sterilized jar, you’re ready to clone.

Cloning your fruits

I’ll skip the part where I tell you to iso gloves and wear a mask etc.  to me those things are a given

1. This part happens very quickly.  Gather your materials on a clean surface.. you may want to work in a SAB, I just make sure there’s no moving air and do it in open air.  I have a very clean house and feel comfortable doing that but your situation may be different.  I like to hit the jar against my hand towards the opposite side of the SHIP.. this moves grains away from the SHIP area so that, A. The grains aren’t in the way of the needle and you don’t stab into a kernel plugging it up and B. It allows the myc plug to get to the bottom of the jar where it will be safe from drying out

2. Before anything else, I use a knife to cut a slit in the foil right above the zip tie in front of the SHIP.  This allows the foil to tear away very easily when I pull the tab.

2. I cut the cap off most of the stem.  I leave a couple inches of stem attached so I can pull it apart towards the cap to reveal the inner cap tissue.  You can also just break off one side of the cap.. however you do it, you just need to expose a decent amount of tissue to poke the needle into, but don’t do that quite yet.

3. Flame your modified needle red hot and set it down.

4. Now you can tear the cap in half and expose the inner tissue.

5. Your needle should still be hot, so squirt a little water out to cool it

6. Now poke the inner cap tissue to get the plug. You may have to poke a couple times to get enough tissue.  If you quickly look down the barrel, you can see whether there’s a plug or not

7. Grab that tab and pull the foil away from the SHIP and quickly inject your myc plug into the jar.

That’s it!  Clone done



With the modified needle, you can use pretty small pieces of cap.  The piece in the bottom is the one I used



If you want to do another clone, just cut the foil on the next jar, reflame the needle and your set to do the next one.

Here’s a couple already starting to recover from their myc plugs



Some colonizing after a shake.. I shake them a few times just to make sure I didn’t pick up any contamination during the procedure.  It takes a few days to see any growth and a few more after that to get to a shake point.  If there’s any contamination, that’ll be evident with the recovery.. especially mold



Now I can use these to GLC to qts and extension jars to preserve the clone or I can dry them out to use later.

This has made cloning so much easier for me.  The only problem I have now is that I have too many clones to grow out



Come this fall, I’ll have a bunch of dried out cultures I can immediately get rocking in some qts.  I’m very excited for that and will give me lots to look forward to this summer while I’m taking a break


Edited by fahtster (11/04/23 06:38 AM)


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InvisiblefahtsterM
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Re: Cloning fruits via Josex’s poke (moved) [Re: fahtster] * 1
    #28301322 - 04/30/23 04:26 PM (8 months, 25 days ago)

This thread was moved from the user's journal.

Reason:
Here we go!


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OfflineRoscoeReturnsS
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Re: Cloning fruits via Josex’s poke (moved) [Re: fahtster] * 2
    #28301375 - 04/30/23 05:20 PM (8 months, 25 days ago)

I like this. Thank you.


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Re: Cloning fruits via Josex’s poke (moved) [Re: fahtster] * 1
    #28301388 - 04/30/23 05:27 PM (8 months, 25 days ago)

Loving this. Thank you.


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Re: Cloning fruits via Josex’s poke (moved) [Re: johnukguy] * 1
    #28325437 - 05/18/23 08:25 PM (8 months, 6 days ago)

I bought some 3 inch 14 gauge dispensing needles (blunt tip) to knock up brf cakes. I took one and sharpened the end so it looks like a biopsy punch. Works great, gives you nice fat plugs.


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Re: Cloning fruits via Josex’s poke (moved) [Re: sprocket] * 1
    #28325453 - 05/18/23 08:33 PM (8 months, 6 days ago)

Nice write up! Definitely will be giving this a try! :syringe:


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InvisiblefahtsterM
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Re: Cloning fruits via Josex’s poke (moved) [Re: sprocket] * 1
    #28325455 - 05/18/23 08:33 PM (8 months, 6 days ago)

:headbanger:


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