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The Corroboree

Darklight

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Posts posted by Darklight


  1. Have you just made up your mind your dad dobbed you in and have chosen to believe it? Seriously?

    How can you be sure it wasn't your co-workers who hate you, or your neighbour or someone whose seen your place or a chopper going overhead?

    You sound like a fractious kid, maybe the only way your dad can really make it really clear he cares is by dropping $7k on a ute for you, because obviously dropping round to chat and taking you out for counteries wasn't cutting it. That's pretty fucken sad.

    Maybe he is as confused by the relationship as you

    Maybe if you're worried about the integrity of your relationship with him don't accept any more gifts from him til you've cleared your relationship up

    Maybe drop round and thank him and take him out for a drink. It doesn't have to be for hours, even one drink is a start. Have a go at mending that bridge for ten minutes. Even if you don't like the outcome, give it a go. If it starts to get weird, tell him you were hoping for a better outcome and don't leave angry, even if that's only to preserve your self respect.

    You sound like you need distance, so maybe Cairns is a good idea. Spend that time setting yourself up there before you go so you have a better chance of success, instead of whining about an expensing present and telling everyone on some forum your dad is a dog, when you have no proof of that

    Plan on coming back all grown up, and take him for a counterie when u do

    • Like 3

  2. Spose it depends on the practitioner, like every other sort of treatment. Sorry you seemed to have gotten a useless person first sesh :(

    My first accupuncturist was useless, no effect at all, so I didn't try it again for fifteen years

    Second accupuncturist was a bloke in the dodgiest looking place, but he was really good. I was scarily functional. Saw him til I moved out of SYD

    Third practitioner was pretty meh and kept insisting I felt better when I didn't, fourth was great.The fifth one was the best I ever had

    Make sure they sit with you during a sesh and don't walk out- sounds like you have both serious medical condition and reasons to be suspicious. They should be working to fix that too.

    Have heard good reports about chiro, and the odd horror story from someone who it happened to. Check round for people you know who recommend a good practitioner. I'm still sceptical but I know people who swear by it

    • Like 1

  3. I am certainly flexible with where I want to be. The main thing for me is warmth/humidity. I would like not to have frost, and humidity helps me with my aches and pains.

    Parts of the areas round the back of Nimbin get Melbourne-grade cold over winter, just not for as many of the daylight hours. The higher up your place is, the less cold it will be IME. But then there can be access issues, especially in the wet.

    One more thought about moving- if you aren't skilled at driving on our shithouse roads, you and your partner both do an off-road driving course before you get here and make sure you have appropriate vehicles. It'll make you more confident. The city hatchback doesn't cut it. We have potholes here deeper than those things are tall.

    A lot of our roads are barely wide enough for 2 cars and in some cases both cars need to have the left hand wheels off the tar to pass safely. Too many people are reluctant to do this and a lot of them are newcomers. We have a lot of passive aggressive/ inattentive drivers up here and they are fucken shitheads who are a danger to life and limb.

    If you can't keep pace and find you are driving 20-50km below the speed limit with cars behind you, indicate and pull off the road, there are heaps of safe places to do this if you don't mind not having wheels on the tarmac. No-one minds if you do this and you will earn respect while you learn to handle the horrible bloody roads

    Nothing spoils a good new neighbourhood friendship like finding out they're the people who have been clogging up the middle of the road at 50k/hr in a hundred zone for 10km. These people are rapidly identified and word gets around

    • Like 2

  4. Take access into account when looking at the land prices - if you're going to have to spend $50K to build an all-weather road into the place, is it still a good price?

    This one's a biggie- at times of heavy rainfall you don't want to be crossing swollen creeks you don't know well, and then climbing a kay up a hill on a driveway with the texture of marzipan icing and ruts deeper than your ground clearance for your bike/ car. There are heaps of lovely places out here but a lot have poor access which gets worse during wet season

    Anything you save on your property you will spend on your car over the next 25 years. The roads are shit. May as well know that now.

    Fencing, water and fire protection are your next priorities after access and housing. Set those up before you get romantic and spend dosh on gardens and gables

    If you look at a bush block, talk to the local RFS if it's not peak fire season, they'll help you assess fire risks and make a plan

    Test soil & water (I think DPI do this?) &/or check with local Landcare, Streamwatch, etc groups regarding any issues. Landcare types will usually be able to tell you a bit about the local soil types & plantlife, including obnoxious weeds & pests that you might be up against. The Bureau of Meteorology keeps records you can consult to get an idea about what weather to expect.

    EAL at Southern Cross Uni do soil and water tests for a good price

    Don't be married to the idea of Byron Shire, as folias says. There are lots of wonderful places everywhere north of Coffs and many are way more reasonably priced than Byron/ Tweed shires. Kyogle even has decent coffee these days. All Byron has is traffic.

    Crazy neighbours are a biggie up here ( tho not as bad as it used to be ) and for various definitions of crazy. If you're not buying onto an MO, try not to buy right next to a large one, because more neighbours = higher risk of more crazy

    If you're worried about isolation, once you get to the area simply join something. Local school or hall association, Landcare, fire brigade, market group, garden club, pony club, anything. You'll meet heaps of people really fast. They will find ways for you to contribute. Oh yes they will. There's lots to do.

    Nobody west of Bangalow really cares what you did before you moved here, it's the contribution you make once you land that's important. That frustrates a lot of people who expect to be taken seriously for something hifalutin' they used to do in the city, but it does level the playing field for people who didn't have those opportunities but do make a significant contribution and have better life and social skills.

    Most of the people you will meet in the first 2 years will not be the people you will end up relating to best, so be cautious about going overboard with your early friendships. Getting to know the locals takes time. After a bit you will become one of the locals and that will make more sense than it does now

    • Like 3

  5. I would give someone else's kidney to see that discussion

    Fuck incog, that's my happy thought for today. Pls upload to Youtube if it does happen

    • Like 2

  6. Bad news in this case unfortunately. I have both an L. shimeji saprobic mutant and the wildtype mycorrhizal in my library. Looking at my notes I suspect I have innoculated with the saprobic mutant instead of the mycorrhizal, sigh. The nutrient source was so poor, it was behaving like a mycorrhizal and didn't notice.

    I hate it when that happens. Has happened to me too in the past :/

    DL - It's been too long between chats. I've been in the trenches with lab work, but I'm on the other side with a few good ideas I'd love to chat about :)

    Yes please, would be lovely to catch up. I'll SMS you tonight


  7. Not necessarily- I've found sometimes a small fruiting form from the original mycelial inoc point when the new substrate is inappropriate.

    Is like species goes into panic mode cos it can't send out vegetative growth too well, so opts for sending out spores.

    But I guess we aren't talking about the same species :)

    I hope you got a win!


  8. Yes by true colonisation I am referring to active production of carbohyrases and lignin peroxidases and active growth. What I suspect is happening is that the fungi are merely savaging free dissolved sugars and simple polysaccharides for exploratory growth, or still using the agar plug. My point is that is difficult to tell from a morphologically, whether the substrate is being metabolised or not... at least at this point in time.

    I've experienced this in experiments as well.

    Best thing you can do is wait, especially if it's a small plug + a large amount of substrate. The inoculated plug will lose it's resources and the mycelium won't find food elsewhere

    You can also run the experiment backwards just to be thorough- add some identical sterile substrate to a colonised agar plate and see if the growth pattern is the same

    Also run controls, just in case there is something weird with the substrate. It happens.

    • Like 1

  9. Yup. Bulk up on household stuff if you can see a hard time coming too

    Sucks to run out of shampoo, conditioner, toothpaste, bog rolls, cleaning products and laundry detergent when you're trying to look good job hunting. Especially if a few of them run out at once

    All that suff lasts a long time. If you know the next few months will be unwaged or hard, go to a cleaning products place and stock up

    • Like 3

  10. I use a $20usd (shipped) blue plastic probe from Aliexpress, this is wired up to a simple pH circuit and fed into an ADC and to an Arduino monitored via the serial out

    This sits in a saltwater frag tank all day and the data gets logged to a xively feed; when the lights come on there is a nice shift from the photosynthesis kicking in, and it's all exactly the same as the colorimetric 7 to 8.9 test kit that I have which is highly regarded (sailfert)

    As for probe calibration, Im sure you're aware that 99% of calib is done by cleaning the probe/keeping it clean

    I use a 12 bit ADC and this generates a value between 0-4096 which is directly proportional to pH

    The main calibration factor used is drift, which is the measure of how different the probe's reading is to a "new" probe if that makes sense

    Nice workaround :)

    I might have to risk a probe shipped from China as long as the specs match mine. I do need it now tho and shouldn't risk the shipping lag esp if it doesn't work, but $ is the limiting factor this week

    As for the rest, to set something up similar to that would be more than the cost of a manufacturer's probe from Amazon and take me a few weeks to work out, it's not an option right now as I don't have Arduino capacity

    But yeah, I love all that DIY stuff when I have time


  11. .... and the answer is NAAAAHHH

    Not all BNC connector pH probes will work with all meters

    It could be because the one I borrowed to use on my unit was a combines pH and temperature meter, whereas on my unit the pH and temp are separate probes off separate plugs

    I couldn't calibrate the meter while it was on there, there was too much difference between the electrode reading and the buffer pH for the calibration to be accepted

    And there is no trim on the unit to be able to manually set the pH

    It's a bugger, I need a precise pH reading and no paper strips I have are sufficiently sensitive to give direct and replicable colour changes, even tho I have a pH 4-7 paper set

    Might take it into a pet store and test it with the aquarium pH probes

    Good news is I found a cheaper Hanna genuine probe. Bad news is it's still $130AUD. I don't want another unit, I want this one to work


  12. ... and yeah, great, regulating companies which sell DNA sequences. That's gunna seriously stifle some research down the track. It's so dumb somebody will actually do it too because Think Of The Children

    Let's face it, this is highly unlikely to become a backyarders process within any realistic time frame, right up until the specific yeast ( which I'm not familiar with ) is available in kit form to idiot-proof the process entirely. And anyone benefiting from the workup is most unlikely to be interested in proliferating the cultures via kit, so that's self limiting for a couple of decades ( by which time other teks will be in the ascendancy )

    I haven't seen the paper, but normally ( and keep in mind research changes exponentially every year ) if you mod something there is a selection gene and a marker gene attached to the gene of interest, so that non-transformed cells don't live and outgrow the transformed cells.

    Theoretically that's a major stumbling block for backyarders, the dodgiest of whom will not understand the requirement and whose batches will fail to produce because of it. Or who won't be able to access the selection chemicals. Or who won't keep enough library stock back so their culture changes down the generations

    ( Edit: don't forget I was wrong yesterday in this field, so YMMV )

    Things which are likely to happen before this drugcopalypse:

    Law enforcement panics are perpetrated via the news cycle and suddenly you need End User Declarations and ID to procure anything which could be 'diverted' for anything sciencey, like pressure cookers or dextrose

    Dodgy bogus yeast cultures purporting to be of this strain are sold for $stupid on ebay and distributed on the grey market to people who won't know what they're doing and won't be able to work out why their sludge isn't getting them opiated.

    Law enforcement comes up with a brilliant plan to extort several million taxpayer $ to train three specialists in forensic prokaryote genetics and nobody will call them

    The panic will subside, but the legislation won't be repealed and you'll still need an EUD and a no-knock raid to buy a spatula

    PS I am most cynical. And I haven't read the paper yet

    • Like 3

  13. the Smolke et al plan had an air of underpants-gnomes to it... phase one, create reticuline->morphine yeast; phase two,????; phase three, morphine!

    (*hilarity*) Underpants-gnome. Is that a Linux thing?

    Love the term. I've seen a few cross-discipline papers which use that method, everyone assumes some term and process is understood by the other specialists and the fuckup/ omission/ swiftie escapes scrutiny.

    I'm stealing your underpants gnomes


  14. I have a lovely digital pH meter which has been a rock solid unit for 15 years. It's a corker

    But now the electrode is almost out of juice, and it isn't refillable ( I checked the manual )

    15 years is pretty good lifespan for an electrode ( I calibrate regularly ). But I priced an identical new electrode at $260 from the manufacturer

    pH meter has a BNC connector. There are a shit ton of aftermarket BNC connector pH electrodes on ebay for about $35

    Anyone had any experience with using an aftermarket cheapie electrode attached to their lovely branded desktop pH meter?

    I'll do a side by side with the old electrode and a regular calibrate of course

    But if most people's experiences of cheap aftermarket pH electrodes is bad, I won't even bother waiting the 2 weeks for one to arrive from overseas


  15. IME GFP does not glow in the dark, not to the extent that it is visible with the naked eye, unless there have been some recent changes made.

    If that cactus has been modified with standard GFP, you probably won't see it glowing at all unless you flood it with a particular type of blue light seen through a particular screen of orange

    Chlorophyll successfully transformed with GFP glows *red* under that light regime, and it's only the really transparent cells, like root tip cells, which can be easily seen as glowy under a fluorescent microscope

    So unless you cut the cactus open and viewed a slice of it under a fluorescent microscope with those screens, you'd see fuck all glowy. Might be more visible if it were variegated or albino, no idea. Doubt it but. And you'd still need the blue light to see it

    Just leaving the cactus next to the bed at night in it's pot would give you nada

    OTOH I wonder, if it were transformed with that compound which makes glowy mushrooms glow- now *that* is visible without special visuals. CBF searching... it's a different compound. Can't remember

    And I could be wrong, it's not a field I follow. Teks change all the time and fast ( edit: yes there have been considerable advances in the number of fluorescent proteins available )

    If you were caught with one of these you would be in vast amounts of trouble unless it was specifically licensed for release by the Office of the Gene Technology Regulator. Federal. Expect major headlines, absolutely no mercy, expensive legal bills and complete shitstorms for anyone else trying to import anything at all

    And remember- I could be wrong. Teks change all the time. I have repeated this for emphasis


  16. Any information on autoclaving coconut water vs filtering it/cheap ways to filter sterilise?

    Coconut water which is sold for TC is heated to denature the proteins then filtered, there are teks for it. And for standard TC use people will do this with coconuts they source raw ( there was a coconut water shortage back in the late 90s sometime and labs were preparing their own )

    But people can add whatever they want to TC media and prepare it however they want

    I can't imagine either raw, unprocessed coconut water or commercial TC coconut water making it through any filter which would sterilise it sufficiently to add to a TC media post autoclave ( removing the need to autoclave the CW and preserving some of it's raw qualities ) but perhaps if you added it to media and filtered the whole lot it might make it through the standard filters we use these days. Is the difference between filtering the 100-200ml of coconut water and adding it post-autoclave and filter sterilising the whole litre or more of media

    A solution of sterile gelling agent would need to be added post-filter to solidify as it too would not make it through a filter. You wouldn't need it if you were doing liquid culture, but you'd want really good tek because liquid culture is so prone to contamination and bugs love coconut water

    Interesting thought, but too impractical for use at a home level. I couldn't even do it here cheaply. I can sterilise 10ml of a reasonable solution for about $4 if I can source a syringe filter right for the job

    0.22uM is the standard filter for sterile technique at this point in time, gets out everything but viruses

    All of this is subject to change the minute someone invents something new and cool ( like a super cheap microfilter system which doesn't cost thousands and clog up a lot and take expensive filters ) which subverts the current standard of autoclaving for sterilisation

    Short answer: you wouldn't filter sterilise coconut water now at the concentrations generally used in TC. Coconut water sold for TC has already been filtered, but only to bump out precipitates- it's not sterile and is AFAIK always autoclaved with media


  17. One question I have which I did not put very well earlier is this:

    Say I have induced shoot proliferation at the axial buds at an increased level with a hormone.

    Now, will I get a finished plant quicker, if I let these shoots grow longer attached to the main stem, with the rest of the shoots and the top

    and then separate them later, allow them to recover, grow and root.....

    Or, would I get them further along/faster by separating these newly proliferated shoots from the main plant when they have a few nodes,

    so that they grow into a separate plant on their own, starting as soon as possible, like when they have three nodes perhaps??

    I understand there is a recovery time after they are divided, but they would be divided eventually anyway...

    do I produce plants faster, by leaving the new branching out plants on the main stem,

    or, dividing them sooner, allowing them to recover and grow separately?

    Took me a while to work out what you meant, thanks for clarification

    This is a *suck it and see* question, individual to each species, media, growing conditions etc. These are the questions rarely specifically answered in scientific publications and need to be tailored by each lab for their own requirements.

    You could do a side-by-side experiment to see what works for you, but it would only be an important consideration if you had, say, a commercial contract for replication with a definitive and punitive time line that was, say, six- eight months away ( to give you time to run the experiment and change replication protocols )

    One other thing I have been wondering about...

    Is there some way to contaminate plants send out to market, in agar

    so they will be fine if 'deflasked' but cannot be used to produce functionally sterile cultures from/ would be contaminated 'in vitro'?

    Hee hee :) There was a lot of speculation on this in the late 90s- ways to maintain IP, mask TC media so plants were still saleable but the media wasn't suitable for analysis ( if it ever was )- addition of food colouring etc. Not sure where it got to, didn't keep up with it

    Not sure about your functional contam to maintain IP and commercial advantage. Short answer- I wouldn't try it. If there is a fart in an elevator's chance that whatever contam was in there could harm a worker ( spores, people with lower immune systems ) or an ecosystem ( including agriculture ) or a business I wouldn't touch it with a pole with a condom on the end. Nice theory tho

    If you have developed a distinct variety you want to maintain you could patent it and protect that patent by having the plant fully sequenced ( sequencing costs about $1k last I checked but prices dropping all the time ). Patenting is a few $k too, and last I checked a patent needed to be applied for in every country the plant is sold.Then you have to enforce the patent by maintaining intel on whatever it is coming out on the market, a full time job for someone

    I am in favour of patents on plant varieties which are very distinct and have been developed by the patent owner- particularly ornamental species. Some scummy companies were trying it on for older strains of food plants as a kind of astroturfing, which was a bastard act of the lowest kind. Patenting is an ethical issue so YMMV

    Thanks mate for keeping us up to date on your progress, you have raised many many good scientific and ethical questions which are relevant for both beginners and experienced people alike, and you're also helping my professional development as well :D

    I did check the TC email list too, on your advice, and ended up signing on again, the volume there has definitely shrunk since I was a member last time, it's down from about 40 messages a day to 4-5 a week. Most manageable, and the search function on the archives is still pretty OK


  18. Thanks again!

    I will post more/ask more soon, but wanted to reply to a question asked earler.

    Here are some bad photos of the rooting of micropropagated M. Speciosa plants obtained by adding honey and coconut water to the medium.

    Honey was from the grocery store, said to be pure raw honey but who knows?

    Sry for delay, I had to do a bit of background checking to confirm my memory was correct

    How much honey/ how much coconut water did you use per litre of media?

    Raw honey would be interesting, tho I suppose the autoclaving would knock the enzyme activity out as much as it would processed honey.

    All the reading I have done has suggested coconut water has a stronger cytokinin activity than auxin.

    Now the important factor with hormone interactions is not just how much of what hormone in which species, but also the ratio between the hormones you add in combination. So maybe the interplay is important with the coconut water. Or maybe it's some other aspect of coconut water which is beneficial with rooting. About to give it a go in some recalcitrant species, may have more info in a month or so

    Those biological indeterminates can be lovely but so elusive to pin down, but shonman your experience shows that addition may have some benefits for rooting as well

    Inspired me, so I'll try it anyhow :)

    Don't worry about the quality of your pics, they look pretty bloody good to me. Getting publication quality TC pics is super hard, carn, we all know that. Yours are fine!


  19. Mate you totally have the touch when it comes to easy teks and getting good fruit out of this place

    This tek has never worked for me at my place, I have too many fungus gnats, but I'm glad it works for you

    But you put a lot of work into it too and you deserve every success

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