Okay, Okay, Okay it's been a minute since I did a log if a minute is 3 months long but I made some progress. For every problem I solve I discover 10 more ( ͡╥ ͜ʖ ͡╥)
So I want to pick up where I left off
I got all the things I was talking about in my last all those tools to grow E Coli but now I need a sterile environment to do my work in as i've found my technique and disgusting garage can't be solved used a open flame sadly so i've made some drawing for other options i've summed it up to laminar flow hood (shown below),
Biosafety cabinet (shown below)
Or a positive pressure room (shown below)
Naturally being broke all of these would be at least a little shit ᕕ( ᐛ )ᕗ
And that's an important factor that needs to be taken into consideration I have an idea that could work and be applied to all of these if i build the filtration system separately and use standard hvac equipment to hook it up I could upgrade with time whenever I want to upgrade
The plan is a plywood box with a filter and a fan on one end and an hvac output on the other, I'm going to design it to use a higher quality hepa filter and a high pressure fan to pull the air required for some of the larger designs. Also this design makes the whole thing modular and WAY easier to repair that in a AIO design
I have some spare wood, and might be able to shrink the design even further if I use a duct fan ( shown below) so I could hook it up directly instead of having to buy an output for the box.
this should work and shouldn't be too expensive as duct fans are ~$15 and the filter I have in mind is ~$25, so 40 for a filter well hell that's a good deal. I probably could go cheaper but really don't wanna skimp out on the filter as it's so important to the performance of this thing.
I also found a new incubator to use instead of the crappy chicken incubator I found an old food warmer (shown below) with adjustable temperature and ironically enough it works really well for bacteria so that was definitely a win for sure.
Now this is my new immediate goal because if i try to bite more than I can chew I choke for like a month and 1/2 before guilt tripping myself to get back to work. so yup i'm not gonna do too much.
One thing I also wanted to restart was the youtube channel. I actually have some interesting shit I could film and I built a new workbench so I have a place to film it too. it would be fun and interesting and who knows? I definitely wanna come back from my hiatus from hackaday and get back to releasing these logs so that's for sure going to happen.
I'll try to have the filter built by the next project log if i'm lucky. It might be after that because I think I'm gonna try to record the video making and post it before christmas. that's the goal but ¯\_(ツ)_/¯ who knows something might come up along the way but I'm gonna try really hard to get it done by then.
I'll be honest I only have 1 new year resolution and that goal was to do a project for every week in 2024 making 52 projects by the end of the year. and if the channel is coming back I might as well record and edit all the projects while I'm at it.
Now money,
I'm broke at it, I gotta admit. I might do tissue culture or grow plants as I already have to in later steps and the added income would be very nice indeed but it's an interesting challenge for now figuring out how to afford anything. I think i'm going to read plants from test tubes in my free time. I found this nice Monstera Obliqua on Ebay and I might buy some throw them in TC and secure a nice tidy profit down the line. TC and plants is definitely something I plan on doing for money soon and in general regardless
I would really love to start making Diy Bio kits (example shown below), like very good high quality ones that were affordable so people could get their hands on them.
like imagine if everyone had the power to make microfluidic chips with their kids or as a science fair experiment. I would love to run a business like that I'm just not sure what to do. I know I wanna market it towards the k-12 market kind of like the https://amino.bio/ kits. I also know I want them to be cheap as well so that more people can get their hands on this stuff If someone is reading this and wants to help me brainstorm PLEASE dm me because I need it. Anyways that might be a fun side project for later who knows.
That's it for now I'm gonna get busy and build that filter and keep thinking about ways to get money. I might record the process and post it but I will definitely write a log about it regardless hopefully soon but I don't wanna do it too often and not have anything to write in these logs.
go over to hackaday to catch up with plan have somthing in the works
total
$607.66
Okay so I just need something to sanitize them something to grow it in and something to freeze them in
Just those steps
I'm going to make a new list for the tools required just to do those
Pipette kits
Autoclave
inoculating loop
bunsen burner
petri dishes
incubator
LB Broth
agar
glycerol
So from this list I have the Incubator and the petri dishes
Of course I will be needing other reagents but those are the most important
The plan is to use a bunsen burner to generate a aseptic area for culturing cells in the beginning just for startup but
But I gotta stop procrastinating I don't know why but my brain just want to push it away as hard as it can no matter what I wish I knew why
Now for the reagent LB broth is Extremely cheap to mix your own and glycerol is also extremely cheap
The biggest cost here is the autoclave and micropipettes I need to get those down as those will be important now and In the future
I need to get something done
Also I found a new discovery while fishing for treasure in the garage a old food heater the kind that was meant for keeping things warm for long periods of time and it could possible be used as a new incubator
It would work and is made out of metal and glass so it would look good too but who am I kidding I need to stop wasting my time and getting something done already
School has just been sucking up all of my time
Right now I need help picking an autoclave this is very important as having an aseptic working area and technique is key to making any of this work
I need help right now in picking an Autoclave as It will be key I might just settle for a pressure cooker right now and spend a little more money later on for a slightly nicer one once i've made some friends
So I made a small change more reflective of what I want to do
The truth is that my goal from the beginning was to modify plants for the good of humanity
So gotta get started somehow
I just added a few small plant related things
It's all stuff i was planning to do anyways
So this is fun gave myself more work yay
anyways let's see what to talk about
I got the heat block It work I tested it and everything
The new vid should be out in a couple hours if i'm lucky
My audio is bad so I need to find a audio closet or smth
It will get better with time trust me
I think my goals right now are to
1.grow and freeze E.coli
2.express a gene in E.coli
3.tissue culture a plant
4.either insert genes in plant or express gene in bacteria
And then the sky's the limit
I have so much I gotta do at the same time
I'm still clearing out the garage but I have made major progress , I'm nearly finished
Still thinking on the air purifier I'm getting closer to it day by day
But i can't clean a nasty f****** garage with a air purifier with so much junk in it but we will stay working
Also i've been reading some papers and think its time i reveal my plan
The end END goal is to genetically engineer amaranth so it is easier to grow brings higher yields and is richer in vitamins and minerals so i can send it off to refugee camps and cure the presence of anemia
So how will we get to that point
Well first I need to get all the equipment collected and finished
Once that is finished I need grow some plant in culture just to get my hands in it
Then we need to figure out the hormone rations for amaranth which the paperwork on is non existent but i found a old protocol for spinach which is related to amaranth so we will start there
After which its just the trial and error of designing plasmids and putting them into the plant
That's the plan for now it will probably change in the future at some point
I would be extremely thankful for any kind of contribution
Now back to the new plan
From the last time this was seen it has grown in complexity a good deed but they are all needed regardless
Now If i don't attack these problems in a direct line don't be mad opportunities come and go
So har i have begun my collection of consumables and was able to successfully acquire and incubator 100% free of charge
It was an old chicken egg incubator that bleached the living H*** out of
It was a little giant incubator
Looks like this expect more crusty
II tested it and it can hold a temp so for now it should work
Also I cleaned and bleached the fridge so it should be good for use
I've also installed pegboard in the garage and am organizing my tools as we speak im about 90% of the way finished it just takes a while waiting on the 3d printer
The autoclave is just a $50 pressure cooker and might as well be marked as finished
The centrifuges and Micropipettes are going to be gifts
I already have a freezer and and ice bath is as simple as a styrofoam box of frozen peas
The heat block is also incredibly simple just a peltier and a thermocouple
The pcr machine is going to be intresting
I can either snag one on ebay, build one, or buy one of the mini pcr machine kits
The gel box setup is also incredibly simple ~50v power supply and some wires and could honestly be made from the dollar store
A gene gun is pretty advance but their are some good kits and guide to making your own attempting it will be fun but worst case I just switch to agrobacterium
After some research I've decided on a positive pressure room instead of a biosafety cabinet, I don't plan on working with anything particularly dangerous and frankly the biosafety cabinet will cost more and give me a smaller working area overall.
If I need 100% sterilization for certain projects I can fashion up a small laminar flow hood to give me a working area if contamination is a persistent problem.
Also I'm doing research on the pros and cons of different filters and filter materials and have made a plan, the plan includes a multistep filtering process with a UVC bulb for when I need to sanitize the entire room. For now I'm most likely going to be using a multi step carbon fiber and hepa filter is a system similarly to the following infographic
The concept seems simple enough just pull air through tiny holes but as all projects go it's bound to hit a hiccup I can't quite see yet.
I plan on making the room out of a wooden frame with plastic sheets as walls. Will it look like a meth lab, yes but I doubt my neighbors care enough to call the police.
The plan is to turn a third of the garage into a sterile lab, half of it into a workspace, and whatever’s left can just be a relaxed space for me and and friends(nonexistent) or family to just sit down and draw schematics or program.
I’m going to start seeing how much the lab will all cost in total. I don’t think it should cost too much after all it’s just wood, plastic, filters, and a fan.
I haven’t stopped looking for what I could use the space for I but before I reach artificial life and the touch of my finger tips. Adding a couple intermediary steps would be a good idea. Such as turning fibroblast into pluripotent stem cells.
That would make it way cheaper to experiment and take some of the pressure off of each run, and know how biology is I will be needing as many chances as I can get.
That’s about it for for now I have things to do, peace.
So I have a firm belief progress in the world in inevitable, yeah so my sister got hospitalized there's a 89.74% chance she'll be fine so I'm unbothered. I haven't felt I needed to stop my research.
I went to the garage and am getting ready to sell everything I made a list and double checked it with my dad and am very excited to begin finally transforming the place.
I'm thinking about transforming an entire part of the garage into a positive pressure room that will be kept 100% sterile for lab work and the rest of the garage being turned into a space for incubators fridges and machichines and a general crafts and storage space. but from what I've seen from other DIYbio crowd online is that contamination isn't as big a deal as I was afraid of but I would still rather be safe than sorry.
I wish the DIYbio crowd was bigger and more info on this stuff was available. I know their are groups but I don't have any near me. and I can find any places online where they congregate. I wish they had teamspeak or discord servers that had easy access because I could really use it right now..
I don't wanna go overboard on the equipment because I still need to buy all the reagents and media I'll use. God biology is expensive lmao.
I might setup a LLC and make a Thermo-Fisher account.
I've been working on the garage (it's a big garage) and clearing it out to make space for the lab. My dad's a hoarder which for me is a good and bad thing because it gives more stuff to sell, but it gives more trash to deal with. He's been trying to clear it out for some time and was happy I took the initiative. He also said that I could have all the money from selling the literal Thousands of dollars of equipment that was in the garage and basement, so long as I dealt with it and took away the headache. so yay?
While clearing the garage I've also been looking for tools, parts, and equipment I could use I found an air compressor if anyone has any idea of what I could do with that. I also lucked out big and found a brand new looking fridge. I think it was meant for drinks and soda cans and it has a glass door, but the racks are removable and the temperature adjustable so it's a great place for me to store my media and reagents. I also found a fan (It kinda looks like this)
I think it's meant for drying paint, it's extremely dirty. But it is compact with decent airflow so I might tear it apart clean and bleach the living crap out of it and use the fan for the Biosafety cabinet.
I'm also thinking about just making a positive pressure room instead of a Biosafety cabinet I'm looking into the pros and cons of each and i'll see what i come up with after a little bit of research.
I've been looking into what I could use this lab for and found some inspo. I think it would be really cool if I could mineralize osteoblasts and Heart muscles and make a little fish that could rhythmically move around in media, sounds pretty cool but I 100% got the idea from the thought emporium I gotta admit. But it really doesn't seem that hard in essence it's just a mini pacemaker to drive the muscles and something rigid you could grow them on (like a bone). I think if you grew them in a 2 step process where you grow bone cells on a structure and then mineralize them, after which you grow the muscle cells on another graft and put the 2 pieces together. But the media for each of the cells is specialized so i'm not sure how you could work around that, I'm not too worried that's future me's problem.
SapoRano
I tested it and it can hold a temp so for now it should work 
