chironex

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chironex
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  • Latest update and part of the series! https://www.youtube.com/watch?v=7kJ2o7P8D00
  • Secondary update, it's becoming very obvious why our DNA extraction failed. I mean we knew we were using the wrong kit to start with, but proper insect extraction calls for incubation in lysis buffer overnight at 56c rather than the 4 minutes at rt we did for attempt 1. This is very good news! Means come Tueday we should…
  • Did someone order a big load of fuck all? We think the issue was our terrible DNA extraction. Gonna redo that with the new extraction procedure, then try again once we've confirmed we have lots of genomic DNA to work with. If that fails we'll redesign the primers. And if that fails... well I dunno what we'll do but we'll…
  • The three that worked are all from the same thing just at three different annealing temps. Though the cycle was run with a gradient decreasing in temp each cycle by 1.2 degrees. So in order it's 72, 68, and 65c. But ya paper based didn't work but since neither did column extraction we suspect it's an issue with our lysis…
  • Ok so exciting updates and interesting updates. First the less good which is that of all the DNA extractions I did, they all yielded only very very small amounts of DNA. This is likely because the kit I used was meant for ecoli, so the digestive enzyme in the lysis buffer won't work properly on eukaryotic cells. Gonna…
  • Oh, we also seperatly tried the paper extraction method. That's why we're doing 2 sets of PCR. 1 with the paper method and 1 with the standard column method. May end up retrying both if they dont work. Will keep updating as things progress.
  • Did the extractions last night. Did 3 spider and 3 yeast. The yeast were from some plated yeast on selection media, and then some from our halloween brew. All should contain the donor GFP plasmid so hopefully 1 of the tubes comes back positive and we can use it for PCR. We're running 1 set of PCRs tonight and I'll be…
  • @misslitty we found a paper that claims to be able to extract PCR quality DNA using nothing but a piece of filter paper. We're trying it with both our yeast and a spider and trying the PCR's on the result with proper reagents tomorrow or the day after. If this works we'll be like 1/2 way done and it'll be the worlds…
  • @misslitty To be honest I bought it cause I misread the listing. I thought there was Taq in there too. It was cheap though. But I'm gonna get proper dntps and buffer. Found a cheap supplier so gonna give them a shot. No worries, figured that'd be the case but couldn't hurt to ask. Hehe I've got so much planned for that…
  • @misslitty 1. No cause it was a free kit meant to be reviewed... on video. So... 2. the wax contains the DNTPs. It's called easy start pcr in a tube. It's a mix of all the usual stuff with some wax so that when the wax melts it forms a protective layer and the aqueous stuff separates out and drops to the bottom. I'm really…
  • Another update: https://www.youtube.com/watch?v=LOyBYEks550 Tl:dr, The Odin sent me water instead of DNA. Going to have to extract some from the preengineered yeast.
  • Ya but since it's impossible to synthesize the gene, there's nothing I can really do about it unless I build the gene from individual silk monomers which is super tedious
  • Maybe, but look at those gels. There's supposed to be ~20ng and 100ng in both of the GFP lanes. Those should be the brightest thing on that gel since each band of ladder is about 27.5ng. I'm gonna test the donor DNA plasmid solution with a nanodrop to make sure there's actually DNA in there. Assuming it is we can rule that…
  • Update number 1: https://youtube.com/watch?v=8mnj5evjMX4 Ran our first PCR reaction and Gel and something seemed weird. No pcr product was generated, but stranger still the lane that was supposed to have the donor DNA in it was empty in both runs. Makes me thing that the vial of donor DNA that I got from the odin is just…
  • Couple weeks. Last of the supplies should arrive this week, then we'll start on the cloning and plsamid assembly. Should be ready to attempt transfection shortly thereafter. First things first is making sure the species of spider I receive is the same as the gene sequenece I have. If not gotta identify and modify/reorder 1…
  • Oh I'm sure they will haha. Already got a list of potential roadblocks and working on solutions in case we hit them. Ought to be a fun project though
  • So tl:dr, you saw the odins stuff and thought it'd be ok to cook up your own experiment? Nope. Hard no. Absolutely not. As much as I like many of the products the odin carries, the frog one I absolutely cannot stand. And for this exact reason. Since he put that out, there's now dozens of people who think it's ok to play…
  • First, don't tell me to chill. Second, got you one of these "." Consider it a gift, use it occasionally. And no, I don't see where you're going. I'll try and address that clusterfuck in order, but it's all 1 sentence so it's hard to tell where one idea stops and another begins. Like an ouroboros had a baby with a wide eyed…
  • Yup! Got lots more bio videos coming up. Just waiting on stuff in the mail
  • Why exactly do you want to do that? Animal experimentation isn't something that should be done for shits and giggles, especially messing with genes that could very easily massively deform the animal. Personally, I'm not gonna give you instructions for animal abuse especially if you are just doing it for aesthetics.
  • couple things. 1, you can't do bioluminescence in mammalian cells if you're using any of the bacterial pathways. some of the compounds in the synthesis pathway for bacterial luceferin resemble molecules that normally are only present inside mitochondria. If you body sees them, it thinks your mitochondria are exploding and…
  • ya looking at a way to do this without the need of viruses and think we found a solution. it'll be a slow burner though till we can test it. busy with other projects at the moment. Glad you liked the video! Got lots more hard core bio stuff coming up, primarily making spider silk grow in yeast amongst other bits and bobs
  • Oh for fucks sake. If you don't know what you're doing, you shouldn't even consider using crispr on yourself. Hell even if you do, you probably still shouldn't. The professionals that DO know what they're doing are the first to tell you not to do that. Crispr is brand new tech and every week more papers come out showing we…
  • not really. Lemme clear this up so we can move past this cause it's getting real old. He was a weird fucking guy, with horrible health habits. The entire time I knew hew, the extent of the food he ate was a single 3 inch by half inch tube of walnut paste and he always smelled like he was dying. Yes that has a smell, and…
  • There are plenty of good coatings. Most here just like grunge and don't want to pay someone to do it for you properly and want to diy it, in which case, ya there are only so many that can be diy'd. But if you contact a manufacturer, most are happy to coat things in whatever you want. Just make sure it's double or triple…
  • Graphene would make a terrible coating for an implant actually. Getting a perfect layer that's required on anything is nearly impossible with current tech. And even then, it'll do essentially no good because it's a single atom thick. Strong or not, nothing that's 1 atom thick is very strong when met with macroscopic…
  • I've made several videos on the process. The decellularization steps are super easy. It's reseeding it with cells that's the hard part because the equipment is expensive. https://www.youtube.com/watch?v=ddGR7YKuN-I
  • So your big breakthrough is that you're not using replication deficient viruses and you're hoping you can control it? Am i understanding that right? All that'll do, assuming it doesn't just make you sick, is make for a large immune response and the gene you tried to deliver will get chewed out by your immune system,…
  • Ya it's meant to be a one off. And I dunno, almost 4 billion people have lactose intolerance. Even at 100 bucks a pop that's good money. But it'll be a very long time before that can happen. Much more testing needs to be done and it needs to be improved a lot