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The Corroboree

Darklight

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Posts posted by Darklight


  1. 17 hours ago, Tripyamine said:

    Hmmmmm I have a nice starter if anyone wants it le PM me 

     

    I really like bags to grow in for a small scale, it's very popular in the industry too. 

    If you get a bad infection/contamination you can burn your bags and reroll otherwise you can just keep reusing them.

    We use Polypropylene bag rolls and seal them ourselves to the size we need, and then sterilize with 70% ethanol and UV

     

    It's a good rule of thumb to always do a 1:5 inoculation with any algae (1:10 if you can't help it) 

     

    I'll grow up a 5mll culture from a stock or a plate, scale that up to 25ml, then to 100ml, then 500ml, then 2.5lt and then start a bag culture with 10L and gradually increase the volume to 20L.

    Then its too a big 100-200L bag like the one pictured and then to ponds/pools all the way up to 400kL

     

     

    arthrospira (spirulina) likes to grow under basic conditions up to like a pH of 10 so lots of media reflect this as its useful for keeping lots of the contam/bugs out.

    Some good info on making the standard media here but you might struggle with some of the chems, also good culture info too. 

    http://sagdb.uni-goettingen.de/culture_media/02 Spirulina Medium.pdf

     

    Another good media to use that you can purchase heaps of for cheap as is Cell-Hi JW (should be around $40 for a kg that will make close to 1000L), you will have to google around to find a supplier as we make our own and haven't had to buy for a while. 

     

    At the end of the day if you scale up slowly and use appropriate media it's pretty easy to do, Spira harvests very easy compared to some other alga, but chitosan always makes it better! 

    Sparge with lots of air and give em plenty of light (outdoors is better than LED but cheap eBay grow LEDs will work fine just gets expensive >100L)

     

     

     

     

    Wow. Just wow. Awesome, thank you so much! I knew nothing of these.

     

    What's the yield like for a domestic setup? Either per kg or how many single portion meals or meal additions you'd get per bag?

     

    This is now one of my one-day-I'll-get-to-it projects. Glad you got there first :D


  2. That. Was. Beautiful.

     

    Thanks to Ronny, Obtuse and Adele and the EGA crew for putting this together. Was very much worth the trip to Melbs.

     

    Panel was gorgeous, hell, the audience was gorgeous. A lovely mixed bunch of age and backgrounds, some were familiar with EGA's work, others knew more about the myco-side but quite a few knew of both.

     

    Audience had a pretty good grounding in all things mushroomy, which made for some excellent questions to the panel. The book table near the ticket point was often crowded with people who had at least one of the references if not more before they even got there. The venue was awesome

     

    The panel. I am such a myco-groupie. They all made themselves available for chats before the event and during intermission and a few stuck round afterwards and they were all fucken lovely people with a grace I can only envy. Question time was professional, comprehensive and respectful and thus also awesome

     

    If you get a chance, go to one of Dr Alison Pouliot's workshops or forays. She's got a really impressive schedule the next few months, quite a few covering specific species of interest ( how cool's that ) and apparently they book up full- fast. https://alisonpouliot.com/workshops-events/

     

    Sapphire and Fungi4Land: https://fungi4land.com/ about the most worthy book project of the decade. Love her work. I will buy all the t-shirts.

     

    Fungimap and iNaturalist. Best citizen science work on the entire planet ( I will find you that reference Cam, I promise ) Do they even need a link? Everyone knows em. Here, have the link anyhow https://www.inaturalist.org/projects/fungimap-australia

     

    Ah yeah, the film. Didn't get to see it. Sure it was fantastic tho, everyone seemed to like it :D

     

    Kudos to you all the lot of you. And thanks again

     

    Bonus points to me for being lucky enough to hang out with Obtuse heaps, and for working out how to use the bloody coffee machine thing at the flat

    • Like 2

  3. On 30/12/2019 at 4:14 PM, Calaverite said:

    Darklight how difficult is the tissue culture process?

     

    From seed? Easy. A great beginner's project, really worth a go.

     

    Sow a lot, some in individual containers, some with a few per container.

     

    Split the batch in half and put one of the halves in the fridge for 3-4 weeks then return it to your light rack.

     

    If you wanted to start from a live plant it could be more challenging. Start with seed in tissue culture and let us know how you go


  4. On 08/01/2020 at 9:21 PM, RonnySimulacrum said:

     

    The screening will be followed by a mycological panel featuring ecologist and environmental photographer Dr Alison Pouliot, mycologist Caine Barlow, ecologist Sapphire McMullan-Fisher and President of PRISM, Vice President of EGA, research fellow in Medicinal Chemistry at Monash University, and co-lead investigator of the upcoming Melbourne psilocybin trials at St Vincent's Hospital, Dr Martin Williams. The mycological panel will be facilitated by Cam Durnsford the coordinator of Fungimap.

     

     

    Ermahgord. I just saw this.

     

    I am very seriously thinking about heading down to Melbs for the night on the off chance I get to even be in the same room as the panel crew, all of whom are my idols


  5. 12 hours ago, Calaverite said:

    Based in Melbourne I am chasing some quality wasabi variety either division or seed.

     

     

    i sent some down to ReShroomed a few years ago ( mmm, maybe a decade now I think about it ). TC clones of seed grown. They grew down at Colac like corkers and the leaf size was 6x what I got in NNSW within weeks

     

    ReShroomed passed away a few years ago :( No chance of getting plants out of him now either...

     

    allrareherbs.com.au site says they're delivering Wasabia offsets in April

     

    But try the seed too. Even if you don't end up with a commercial-grade wasabi root, the leaves are great for winter salads and the stalks apparently are useful too

     

    Give it a go I reckon


  6. 36 minutes ago, Halcyon Daze said:

    Saved me and my neighbors so far, but some nearby homes have been lost. Even looting has been taking place in some areas, so it's a very sad situation for some. Pretty soon we'll all have to join them on the allied front, I can't see the situation getting any better in the coming decades.

     

     

    It is all so unbelievably sad I'm short of words. I hope you stay OK, and I hope it rains

     

    • Like 2

  7. 2 hours ago, niggles said:

    I took 9 large samples wrapped in paper towel collected with sterile mushroom knife stored in new takeaway containers ready and waiting for you. They come with gps coordinates time and date and photos of original fruiting bodies with a foot for scale. 

    Let me know what you want me to do with it :)

     

    Ooh er, thank you, I'll SMS you. I fell asleep and didn't do those plates yet, but they won't take long tomorrow

     

    I can't wait, this'll be awesome if we get it


  8. 2 hours ago, teamwhy said:

    laccocephalum hartmannii or a form of is my guess

     

     

    Nice :)

     

    2 hours ago, teamwhy said:

    here is a close up of the inside

     

    croppy.thumb.jpg.e2d3ca504c643f23d7d2846cc3496753.jpg

     

     

     

    Is that the inside of the pseudo-sclerotia or the fruit? I'm guessing the former. If so, it doesn't look very appetising, might be a good one to inoculate onto seedlings for re-veg work

     

    What are you up to tomorrow? I'll pour these plates out tonight. And try not to pour too many different agar types, we all know how that ends up

     

    If the inside of the fruit looks easier to culture and breaks up/ cuts well I'd love to grab a fresh fruit from either you or Niggles or both

     

    I'll bring some fresh H2O2 MEA plates for you both too. Best to have 'em fresh before the H2O2 goes off. Never know what you'll find in a hot forest :)


  9. 1 hour ago, teamwhy said:

    Sorry @Darklight truffle was probably a little bit misleading I should have said pseudo-sclerotia. I will post a picture later today. they are still pretty cool

     

    Is alright, I knew what you meant and I was into using the wrong term as well.

     

    Am most interested in getting a tissue sample onto fresh H2O2 plates ASAP. Cancelled all other appointments today, making plates now.

     

    Niggles is hunting samples as well. What are the chances of me getting some tissue samples off you late sarvie/ do you want me to bring over some fresh H2O2 plates?

     

    What tree species are the pseudo-sclerotia originating from? I might have to make up some woodchip media for it

     

    I'm so easily distracted....


  10. 10 minutes ago, teamwhy said:

    check out these guys that are popping up everywhere after the fire. 

    crop.thumb.jpg.da6183a89a13cde9c05f8ad66e7e4332.jpg

     

    You reckon they're the same as Niggles is finding? Check thread

    http://www.shaman-australis.com/forum/index.php?/topic/46677-found-growing-in-the-ash-from-still-burning-bushfires/

     

    10 minutes ago, teamwhy said:

     

    I will go see if I can dig up a truffle this week some time.  I so want to eat one they smell so good! 

    Probably some Laccocephalum spp.  but can find a picture of any that fit. 

     

     

    WHAT? THERE ARE TRUFFLES? YOU'RE FINDING TRUFFLES?

     

    @teamwhy Get tissue samples and dry em proper, at least 200mg. I'll add them to the collection of things that need sequencing ( special kudos to Obtuse, who explained at length why this should be done properly )

     

    Put some fresh tissue down on H2O2 MEA agar plates. If you don't have any I will make some of the bastards up. Niggles might have access to some too, but they will be a coupla months old

     

    *sigh* you prolly don't have time, but if we can grab some properly identified edible native truffles we will all be well stoked, and we can try to add some to any regen work.

     

    Anyone else got any favourite firies out there?

    • Like 3

  11. 1 hour ago, niggles said:

    So much gratitude and respect for anyone who helped in any way. Volunteers literally saved our entire community. Coming to the aid of firies when we had collapsed from exhaustion and taking up the fight, keeping the fire from getting away from us. I've never seen community pull together like that. Proud to be part of it.

    I'd like to share some pictures but please don't post them elsewhere as it may make some official people grumpy.

     

     

    Spectacular pics, hey those first ones aren't Hill 3 are they?

     

    Everyone who's never seen a close up bushfire please note that almost the only time you get a chance to take your phone out and take pics is when there isn't anything major on. For pics you need to take your gloves off and not be holding anything with water dripping out of the end. And not running anywhere. Or yelling for backup on the scratchy radios.

     

    Also consider the stuff out of shot. Like the trees already 90% burning you can't see in the pic 10m back which may or may not fall over and send embers everywhere.

    • Like 2

  12. 53 minutes ago, niggles said:

    Can anyone ID this one? They were some about the diameter or a tennis ball up to nearly dinner plate size! The fire wis literally still smouldering metres away and they were fruiting.

     

    I'm still excited about this one but have done fuckall about it, sorry.

     

    Is it still growing?

     

    Is there any chance our Sorcerer's Apprentice and Mistress of Cultures could grab a tissue sample and/ or a coupla H2O2 plates? And a spore print?

     

    Anything that comes up in the middle of a fireground so early is prolly a pioneer species, could be environmentally interesting

     

    Keep an eye out for fire morels after it rains, and edible native truffles too


  13. Well, shit. Worst season I've seen, ever, and I was here for the '93 fires.

     

    There's a few crew affected, either directly or in the path thereof. Prolly more than I know.

     

    But I've seen such grace on the firegrounds which has pushed past my normal misanthropy, and some of that grace has come from some of you.

     

    Shoutout especially to @niggles and @teamwhy, whose continued and protracted presence onsite is making a huge difference. Niggles mate you lost yer fire cherry well and truly this year, you've been doing some long days, done some hard fucken yards and I'm proud of you. Teamwhy, seeing you on comms and knowing you're there day after day makes me smile like a goose.

     

    Also whoever to put those tiny sweet garden mandarins in the snack packs the other day, they were small and hard won in the drought and sweet as fuck and much appreciated

     

    Shoutout to the civvies at Tuntable and Siddha Farm feeding everyone and pushing firebreaks through impossible country and the yoof being all splendid rolling round on ATVs, everyone's hearts swell just seeing them rock a fantastic and major contribution. Anyone who thinks Millennials are snowflakes should see this lot, they're pure gold.

     

    Shoutout to the firies everywhere. Everywhere I go I see trucks from somewhere else.

     

    Shoutout to Mudgee firies who came up to help out at Drake, now they've got their own shit going down locally like a few other brigades who went well out of area up here. And the CFA ppl in VIC, I think we have a couple on here

     

    And none of this Hero Firies shit. Firies are you. This hero worship thing just separates the people who do the work from too many people who can, but aren't. Join up, help out, at the very least clean your fucking gutters, talk to your neighbours and get your evac plan in line ( yes, I do understand that not everyone can kick in and the entry requirements are onerous these days, that's for another thread )

     

    You should see who's out there, it's amazing. The fires are fucked and the environmental devastation is heartbreaking. The humanity so far is splendid

     

    Pray for rain. Dance harder, fuckit

    • Like 9
    • Thanks 1

  14. Right now what I'm doing is drooling. I don't understand 80% of the techspeak, which is entirely normal for a new field for me, so I'm not bothered by that. If I needed something doing in this space right now I'd spend the time because obvs the field is expanding and I need to watch the directions. All I can do at this point is observe, and drool

     

    What excites me is that there are people here with relevant and current skillset who can explain it well and who are sharing information

     

    Would it be possible for 2-3 of you to get together and put something up for the next EGA Garden States? Like a DNA 101 workshop on a single relevant point to us ethno freaks?

     

    A few of us have been working on practical protocols for base-level workshops for the day next May. Light microscopy, a little TLC, and something fungal again. Stuff people want to do at home- and can- but just need a few pointers to get the most out of the tek


  15. On 20/08/2019 at 10:39 AM, Tripyamine said:

    Kinda funny but, there is a legend from ANU that has made a group on a website called protocols.io that has customized extractions people have perfected for heaps of different tissue.

     

     

    If it's the same legend from ANU who was proposing exactly that a few years back, please tell him I said hi? He was a legend even back then, now I reckon he'd be a fucken rockstar

     

    Great site too


  16. I don't understand about 80% of the technical speak of the above, and genetics isn't my field

     

    Am incredibly impressed tho, by how much the skillsets of youse all have progressed :)

     

    It's been nearly 3 years since the OP put up their request. What's changed?

     

    My understanding is that SDN-1 knockouts are now no longer covered by OGTR legislation, but not sure if this covers environmental release as well. Anyone know?

     

    Quite possibly what hasn't changed, and may be going backwards, is public access to a lot of new databases.

     

    And I suspect ( but cannot confirm ) that the human factor is still a big part- quality DNA extraction is hard for some species or plant parts and as Tripyamine says above, a bad result needs to be caught before it proliferates. Add that to the variables of technique and machine and it sounds like it'll be a shitfight for a while yet


  17. On 26/07/2019 at 7:28 PM, Maximt said:

    I think why most mush cultivators use horizontal is because its cost effective. i.e. you can DIY - find a filter, a pump and some ply and away you go, and you can set up a 3-4 meter wall of these for relatively cheap. And if you are doing mass inoculation it makes sense and saves time, you can line up all your bags on the desk, clean them all at once and just go through inoculating.

     

     

    Not IME. Horizontal airflow pushes sterile air towards you so if anything falls off your hands or tools while you're working, it is blown forward, away from any media or tools which might be contaminated by falling spore. Cost effectiveness isn't much to do with your choice, you want horizontal flow so as not to contaminate your work while you're working on it.

     

    Cheap HEPA flow units can be great for most people doing myco work and spending more won't necessarily guarantee success if your technique is poor or your existing unit has issues.

     

     

    On 26/07/2019 at 7:28 PM, Maximt said:

    Vertical flow hoods are generally something that is used in laboratories i medical research facilities. The save space, as the filter and pump don't take up desk space. They are generally enclosed from all sides but 1, and have positive pressure to push air out through the bottom of the table and the 1 opening. But if you need a lot of these, i.e. 3-4 meters, becomes very expensive, especially if they have to be custom built. Added bonuses are they are enclosed from all sides, hence less chance for contams to get in, just 1 opening, and take up less desk space.

     

     

    Nup, not IME. Vertical flows are used where there is a danger to the operator or environment from pathogens, and they're often used- routinely used- in medical/ clinical laboratories where they are testing or researching organisms which can cause disease. Pathology labs, working with blood- or air-borne viruses, Zika, HIV, infected tissue of any kind. Biosecurity labs where they may be testing imported plant tissue for bacteria etc, or extracting DNA/ RNA for further tests.

     

    At least one of the Biosafety cabinet classes does both kinds, for work which is required to be sterile and where the operator and environment both require protection. Air is first pushed through a HEPA filter at the back- then rather than being pushed forwards towards the operator, a second air current near the front of the cabinet works like an air curtain and forces the air downwards before it gets to the operator.

     

    I've worked in one for Biosecurity purposes, it's not my normal cabinet, but they've gotten a lot more specific on classes and subclasses of cabinets these days so the changes over time haven't registered on my radar. I do know I've rarely worked in a Class I cabinet ( fume hood type ) and never in a Class III cabinet ( glove box type )

     

    Happy to be proven wrong, I'm out of touch these days

     

    Read this again: https://www.thermofisher.com/nl/en/home/references/gibco-cell-culture-basics/cell-culture-equipment/laminar-flow-hood.html

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