blender-tek cloning - just an FYI

Started by substrate, March 22, 2010, 10:47:38 AM

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substrate

I had read about the "blender-tek" method of cloning:  cutting an uncontaminated portion of the mushroom stem, putting in blender with clean water, then inoculating the substrate with the juice.

Tried it with oyster mushrooms (picked same day) from the store.  Inoculated six jars pf-tek-style and two 6lb bags of pasteurized wood chips just for kicks.  I've been interested to learn if one can go direct from liquid culture to bulk substrate - if you use enough (a LOT) of LC.

Myc IS growing.  Unfortunately all of the 1/2 pints have trich right at the point of inoculation.  I'm guessing i was not clean enough when cutting the insides of the oyster stems.  jars are almost fully colonized at three weeks - but since they also have trich - they are all going into the compost. 

The trich had to come from the LC itself (unclean piece of stem to create LC) or from the re-used/sterilized syringe.  Thus far the wood chips are not contaminated, so I will let the myc run and see what happens.  If they grow I can assume the trich came from the syringe and not the LC.

So my point - blender-tek works!  You just have to do it cleaner than I did...

pics are hard to make out (crappy phone camera), but you can take a look here if interested:

http://s979.photobucket.com/albums/ae279/alt0182/lolie%20gaggin/?action=view&current=Photo_031810_001.jpg&newest=1
http://s979.photobucket.com/albums/ae279/alt0182/lolie%20gaggin/?action=view&current=Photo_031810_002.jpg&newest=1
http://s979.photobucket.com/albums/ae279/alt0182/lolie%20gaggin/?action=view&current=Photo_031810_003.jpg&newest=1
http://s979.photobucket.com/albums/ae279/alt0182/lolie%20gaggin/?action=view&current=Photo_031810_004.jpg&newest=1

substrate

Quick update:

On that original test run I "inoculated" two 5lb bags of pasteurized wood chips as well, literally dumping the LC on the substrate.  One bag did nothing, and there was a bit of trich (not too sure about my pasteurizing methods).  The other bag actually is about 1/4 colonized with oyster myc.  Go figure.

On my new project, while prepping the mushroom stem for the blender I put one 1/2 pint jar with H2O2 and another with purified/boiled water.  Once inside the glove-box I dropped the piece of stem into the h2o2 and immediately into the water.  As I trimmed away the outer portion of the stem I would dip fingertips and utensils into the jars to try to kill contams I had come in contact with during the trimming process.

I did this with three different pieces, all blended and inoculated into prepped popcorn separately.  Thus far, 11 days in, no contams and myc is well under way with all three. 

I think the h2o2 is a legitimate step to add to the blender-tek cloning process.  It takes some care - hydrogen peroxide will eat your entire mushie in a matter of seconds - so a QUICK dip then an immediate bath in pure water is really important.  I have no doubt that some of the mushroom matter that went into each blend was dead - but there was still plenty of living material for a successful inoculation.

substrate

Almost three weeks in, fully colonized popcorn.  Ready for bulk.

ImAFungi

Awesome man. Can't wait to see some pics. You think they'll be ready in time for this months contest?

           -ImAFungi
String Cheese + Sherwood Forest = Rothbury Incident!!!

substrate

Pasteurized the straw and did the oysters in-a-basket last night. 

If my neighbors didn't think I was nuts before, weed-wacking straw in a rubbermaid tub in the driveway will have an impact (that stuff gets everywhere).  It was like standing in a big golden tornado.

I didn't have quite enough straw prepped to do the Eq basket, so I did one big cubesagna baking pan (straw, popcorn, straw, popcorn, straw - like lasagna) with one qt jar, and will do the full-size basket this weekend. 

I hope to have something to turn in for the MAY photo contest.  In the meantime you're welcome to browse my photobucket:  http://s979.photobucket.com/home/alt0182

substrate

#5
From the original blender-tek cloning attempt...  Two months later, the pile of dead leaves and moldy substrate kicks out over a pound of oysters.  I guess nature finds it's way.

Sliced Ed 'em up and ate them in some chicken broth with green onions. 


The laundry basket tech got pretty bad trich before I could get to full colonizAtion. Tossed the whole basket.

cugak

Let me ask you, when you filled the basket you putt it in the plastic container,  closed it and putt it in the dark for 2-3 weeks to colonize??
share international

substrate

Yeah.  20 days to colonize the popcorn from the blendertek clone LC.  Within 3 days of creating the basket there was myc spreading and growing within the straw.  Just 9 days later saw the first signs of trich.  I'm guessing I blew the sterilization somewhere in the process, possibly on the BASKET itself.

This was with the oysters.  Different story with the others. 

dub504

Substrate, did you pasteurize or sterilize the straw?

substrate

I pasturized the straw, a long process (lot of work) for no more than I got out of it.  I did my best to sterilize everything else along the way.  The basket I used was a hanging file folder from office depot - like an old milk crate, but with bigger holes.  It had a lot of odd angles, and I think I failed to get it really sterile.  Should have sprayed with alcohol.

dub504

Sorry you had a fail. There's always next time...