Darklight

Just thought I'd let you know: I have had a 100% sterile success rate with the peroxide agar kitchen tek, both with the cooked method ( no autoclaving ) and the autoclaved method. Methods and comparison below Dispensing and media transfer was done under absolutely not sterile conditions, in order to see if the tek could be replicated anywhere ( those of you who know my kitchen can stop laughing ). I was a tad sloppy with my transfer tek as well to see what would happen Reporting it here because sometimes the signal/noise ratio can be a little loud elsewhere Batch 1- autoclaved + 6ml/L 3% H2O2 Chinese sauce containers were autoclaved in a bag for 20 min 100ml liquid MEA autoclaved for 20 min, consisting of the following 20g/L light malt extract 0.1g/L garden lime 0.1g/L potassium phosphate dibasic 15g/L gelcarin ( 20g/L agar works just as well ) Media was not subject to a pH test Post autoclave the media was allowed to cool and just above setting point 6ml/L 3% H2O2 was added. Media was swirled heavily so that the inner surfaces of the bottle were fully coated Dispensed immediately into sauce containers *on the kitchen bench* in the open air Lids were placed lightly over the containers and 1hr later the plates were completely sealed after inoculation with various species Batch 2- cooked 1hr + 6ml/L 3% H2O2 Chinese sauce containers were autoclaved in a bag for 20 min 100ml liquid MEA cooked for 1hr by placing media container in an open saucepan. Media container lids were left loose. Water came up the the media level- bottles weren't more than 3/4 immersed. Cooked at a slow boil for 1hr 20g/L light malt extract 0.1g/L garden lime 0.1g/L potassium phosphate dibasic 15g/L gelcarin ( 20g/L agar works just as well ) Media was not subject to a pH test Post autoclave the media was allowed to cool and just above setting point 6ml/L 3% H2O2 was added. Media was swirled heavily so that the inner surfaces of the bottle were fully coated Dispensed immediately into sauce containers *on the kitchen bench* in the open air Lids were placed lightly over the containers and 1hr later the plates were completely sealed after inoculation with various species Sterile ( non-peroxide MEA ) library cultures were opened and haphazardly used ( left open for much of the inoculation process ) to inoculate the plates above using a scalpel blade which was only cleaned and flamed between species The following species were placed in the centre of the agar of each container Reishi ( Ganoderma lucida ) Blue Oyster ( Pleurotus spp ) Lion's mane ( Hericium erinaceus ) Elm Oyster ( Hypsizgus ulmanarius ) Plates were sealed with Austraseal and incubated in the dark at 22C 2 plates from the cooked group and 2 plates from the autoclaved group were left uninoculated as controls to check for contamination during dispensing By my judgement it was all a bit haphazard and I didn't believe it would work. Even a contam rate of 10% per batch would have been acceptable At +1 week there is no contamination, anywhere, and growth is good for the Reishi and Elm Oyster. Still waiting on the Blue Oyster and Lion's Mane, but plenty of time yet- those parent cultures could have been a little old- I have storage/ library cultures and can reinoculate from them easily at +3 weeks If you are thinking about the peroxide tek for agar- give it a go. I've only made it sound complex cos I wanted to write it all up so you know I took all the steps. I now pronounce this part of the tek piss easy Edited very fast, because I am an idiot and forgot to put the decimal point in

Seriously brilliant work Ronny and crew. Outside the lab I am so completely flummoxed by anything more complex than the construction of a toasted sammidge. You all putting this together for the community is so absolutely brilliant I can't even...

Awww.. I am so honoured that you of all ppl Obtuse are squee-ing at the thought of this, I really respect and admire your work Can't wait to do the Myco-agar workshop alongside you- that too will be an honour

Ethnobotanical Research 101- starting from scratch Torsten and I are running a workshop on Laboratory Experiment Design at EGA 2017 We want to share our love of citizen-science. To convince you all to partake in the formal, logical process that can answer so many of the phytochemical/ ethnotoanical questions you've asked over the years. Wonder no more! Act! Workshop's for beginners and wrinkly old lab-hands alike. Anyone, literally- anyone can design a simple, robust protocol which gives solid results and contributes to the sum of human knowledge. Those of you with extensive practical experience in experiment design are very welcome to share the ( sometimes bitter yet hilarious in hindsight ) fruit of your work with us huddled masses. It's not rocket surgery. Lab experiment design is a simple checklist, a bit of planning, some thorough checking and the resilience to simultaneously accept and critique the data as it falls. Carn, we all talk about experiments we'd like to see done. Or exceptions to established practices we've seen work. Shared variants or refinements of new teks. Wanted to know why. Or wondered why the hell something didn't work out after we ( mostly ) followed the instructions. Workshop's interactive. Which means we need your input. Some of which can start here on the forums- reply with some pointers about your experiences or plans. During the workshop we'll welcome your thoughts, interjections, inspirations. Keep 'em coming, keep it moving Workshop's practice-based. Inasmuch as we're pointing at issues around design of theoretical experiments involving the legendary ethnobotanical Dragibus curiosa. Not sure what kind of experiment yet. Help us decide. A simple germination experiment? Optimal fertiliser requirements? The virtues of rhizobial inoulation? A cost/benefit comparison of propagation practices? Testing storage parameters for volatile compounds in the dry product? Determining genetic markers for drought tolerance? We'll settle the best questions on the day It'll be lighthearted. There *will* be lollies. Like all good laboratory-grade successes, some of them may be thrown at you, randomly. Some you must earn. Fate favours the prepared, apparently. It's serious business, experiment design-but that's no excuse not to have fun Bring your questions, your experience, your weird attitudes and your sense of humour.

Most of the things you've ever wondered about can be turned into an experiment so you can get results and stop wondering... Hell yes. Even if your results are inconclusive you get to rule out a bunch of stuff and know where you stand with regards to progress. Every step is a step forward. You often do not need as many of the shiny toys as you think- the best thing you bring is attitude, reliability, accuracy, resilience and resourcefulness. Most people have these. Framing your question so it produces valid data is the next step. You may have noticed that both Darklight and myself focus a lot on fun. I think that's because, for many of us, science is such a social process. It's fun, joyous, revealing, sharing, infuriating, frustrating and above all it's creative. I never learned about those aspects in school ( I was kicked out of HS science for asking too many questions and stayed away til my early 30s ) Yep. Creative. Who'da thunk? I find the varying levels of experiment design, planning and execution similar in interplay to a Bach fugue- yes, the structure is important, but it's the contrapuntal nature of the different aspects of the process and the precision you bring to the work which evokes beauty and joy. Good science readily lends itself to networking, meeting more weird people with similar-but-different viewpoints on so many things who will teach you so much about your field and your worldview. I hear great research ideas every other day. Followed closely by all the reasons why that person isn't doing them. Yep me too. Those convos are so sad. Too many people underestimate their own talent and ability to contribute. Citizen-science projects like Fungimap and Atlas of Living Australia are prime examples of the scientific method being socially productive and fun- as well as making valuable contributions to scientific knowledge

Seriously, youse all need to come to EGA. I do not use the word need lightly in this instance. 90% of the people who would be reminding you why you need to come are fully immersed in actually setting it up. Drowning in it almost, just so you will have a deadset brilliant time I've been on the very outer edges of the org team and I have seen so much legendary epicness coming to the event- so many brilliant presenters and things and food and.... just you wait!

Damn, i thought this thread was going to be about weird things people find in their handbags Must be just my handbag then

Thanks mate, they arrived today. Frantically googling best standard propagation processes right now

Brugmansia. Always Brugmansia. Plant a few different cultivars. Not only do they attract heeeeeaps of bees, but when it's a drizzly day and the foraging's hard work your bees will still work the flowers, hanging out underneath them like umbrellas, waiting for it to be dry enough to fly back to the hive The resultant honey is not hallucinogenic, at least IME

No idea about yr prob, sorry. I'd suspect bacterial, because of increased humidity and reduced air flow, but can't be sure I'm 100%. Is a good lesson to learn- when changing any parameter, keep some material back in it's original place. Always. Not only does it mean you have reserve material, but the remaining group acts as a control so you can compare. I'm not being unsympathetic at all- I recognise the temptation for a quick fix/ progress, and it's one I've succumbed to occasionally myself with the same results :/ Goodonya for staying on top of it and making sure your babies didn't suffer too much for too long

Dammit, I want the scissors. And a Ronny print. Want want want

Thanks mate, am impressed again by your ability to log and share your data. Many thanks!

Aw nice one Amz! I'll donate halfa dozen aseptic cultures of an Acacia, all derived from same parent plant. 100ml tubes/ 30ml media for some keen soul to have a play round with in tissue culture. Deflask 'em straight away if you want, replicate, send to callus or mutate, they'll be all yours

And an absolute fuck-ton of excellent seed Master B's sent me brilliant instructions for handling the albino and others, and a bunch of photos of the mother plants as well This is beyond generous mate. Beyond. Thank you so much!

Unpacked, they revealed these. Not only the expected offspring, but a couple of generous extras. Like, extremely generous The middle one is a Zellys melted wax slab graft, the other outer one is a genuine medium tbm

O M G. Thank you Master B. Parcel arrived super fast, really well packed, babies in excellent condition

IME unless you have super-dry facilities for it, it won't do well. Or even live. At least here, every time I have tried to deflask it, even the ambient moisture kills it. Something in the air maybe, bacterial perhaps, it does fine in TC so far with+90% ambient humidity Herbalistics sometimes has seed. Could be worth a shot that way

Happy and excited and super impressed by the EGA lecture and workshop lineup- final list is here: Entheogenesis 2017 lecture and workshop list Seriously good work from the EGA team, many thanks and congratulations. See you there!

Yea verily, I do know it's for the pictured graft of beauty You're the fulla who did the work. I think you just set a new forum record for stunning behaviour

She's a beaut too

$555 I WIN!! Do I win? Thanks so much Master B, this is an excellent lovely fantastic thing to do

$555 Just in case

$350

Are there any electronics geeks who can tell me why the f*k PAR meters are so expensive? The standard models don't even have a logging function FFS, and start at around AUD $300 PAR ( Photosynthetically Active Radiation ) meters measure light based on that part of the spectrum which is used for photosynthesis by plants. https://en.wikipedia.org/wiki/Photosynthetically_active_radiation Dead handy little buggers those meters. Our human irises open and close to adjust the amount of light they let in, but plants don't have this light-adjusting capacity the same way. So a plant can be getting much less useful light than we realise til we test it. Plants in tissue culture photosynthesise way less than outdoor plants because they use the sugars ( or other carbon sources ) in the media for energy. It's why we get away with using fluro tubes and get good growth. But fluro tubes lose a large proportion of their photosynthetically useful light after 3 months and need replacing- the light at those wavelengths isn't differentiated by our eyeballs, but the plants notice, believe me. I've been told that other indoor lights- metal halides for example- also lose some of their PAR strength after a period of time, but I have no experience with this. And I've seen people swear blind their plants are sick, when after careful checking their plants aren't getting enough light because they're shaded for the parts of the day when there's no-one around. This is more pronounced in winter when the days are shorter and the sun's angle of attack is different I bought plans to make a unit with an Arduino using LEDs, but it's a world of hurt for me and the tek ( to my n00b eye ) seems to be primitive and possibly inaccurate. It'll have to wait until I have some geek mates in-house for a weekend with soldering irons and whisky and those weekends never turn out the way they're planned anyhow :D Fuck it, I want a network of the little buggers with temp and PH, two sensors it's easy to build loggers with. Then I can really see what my plants are up to. Plus it could save me some $$ replacing fluros all year. And maybe save some planty lives if there are dodgy tubes in the rack Anyone know of a cheap chip that's up to the job?

Bugger, forgotten how to delete topics Seems there is a bunch more DIY stuff out there than there was last time I checked, despite the commercial units being the same price I need to do more reading on the DIY stuff and try to evaluate how good the latest teks are for PAR