I Built a Positive Pressure Clean Room for CHEAP

Every good scientist needs a lab, so this week we're going over how I built my new one! This lab is designed for biology work, specifically genetic engineering of bacteria and yeast. That said it can easily be modified to work with plants as well. As a positive pressure room it could almost be used for working on hard drives and other sensitives components if you added an air lock.
This video is the first part of a series where I'll take you through building and stocking a genetic engineering lab.
Follow up video:
My New Lab - • My New Lab is RIDICULOUS
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[@RandallStephens397]

I love the warning about radioactive high-voltage laser-emitting biohazards that make the floor slippery

[@bok..]

banana

[@ChromicQuanta]

@@bok.. Just a reminder that 4y ago, you replied "banana" to a random comment!

[@Lelemon2373]

@@ChromicQuantaust a reminder that you replied to a random comment replying to a random comment saying just a reminder you replied to a random comment saying banana 4 years ago 4 months ago

[@ChromicQuanta]

@@Lelemon2373 Thank you for the reminder!

[@vwlz8637]

​@@ChromicQuantareminder that u need to take out the trash

[@fatkorn]

I'm not a scientist type like you, but I do some small mushrooms work. To keep stuff clean I use a pair of old home-grow tents an my "lab" is a sandblasting cabinet from harbor freight. It's got mounts and plugs and switches already wired for lights, I use a uv and a white light LED. There are fresh air and vacuum hookups that fit hepa filters an such. Works good for agar plates an inoculations. Since I didn't need to buy tents an metro shelving my set up was nominally $200 with one them 20% off coupons.

[@devrim-oguz]

UV lamp will screw the plastic on the walls. It makes them rigid and brittle (at least thats what I think)

[@leobla744]

in his stream he said that he would replace the plastic if neede because its cheap and easy (sry for the bad english)

[@Kalanchoe1]

I agree I think the ozone and UV being generated will make short work of that thin plastic.

[@lorow2542]

It really depens on the type of the plastic used. For example, ASA is quite OK with UV

[@ExtantFrodo2]

Since even newspaper can block UV, shielding it should not be any problem.

[@xWood4000]

I would install an alarm for the UVC lamp, both sound and light.

[@colbiesthename2875]

Not a bad idea, and it would be super simple.

[@Avocado-yw4xb]

hmmm yes

[@DoRC]

Lools good but wouldn't it be better to put the exhaust on the other side of the room so the airflow isnt just looping in that one spot?

[@JK-de1oy]

Interesting idea but air doesent work like that, the whole room has to be pressurised and filled with air for the air to be pushed back out, it isn't sucked out as that would creat a vacum.

[@ExtantFrodo2]

The design is more to keep stuff from the outside getting in than for providing air to breath. That size room has O2 enough for probably 72 hours of work and there's no bed in there so...

[@MsMotron]

i am a physics bachelor working in a biology lab, and i used so much mathematics physics and chemistry there. also are you even allowed to do genetic manipulation experiments?(not that i care) in germany where i work we have rediculus restrictions on any genetic reserch. i work with genetically modified human cells, that will die if you look at them funny, but i still have to make absolutly sure they can not leave the laboratory and contaminate the environment.

[@MsMotron]

jeah i know but the things i am doing or he is talking about in this video dont occur naturally at all. you dont just evolve jellyfish proteins in your retinas.

[@bjarnivalur6330]

I'm really not surprised that the German government is super strict about anything to do with genetics, especially human genetics.

[@MsMotron]

yes but only in single cell organisms. there is a big difference between natural and artificial genetic manipulation. natural usually is a lot slower and more random. even in dna incorporating bacteria. i can understand why one would be careful modifying crops for example. but as i sayed in my case i think it is overkill. Also when your eyes have become flashlights i will be really impressed. one can also say it is natural for species to migrate to remote islands, but that does not mean we must not be careful introducing new species to them.

[@MsMotron]

good one! but these rules are actually imposed by the EU.

[@MsMotron]

dude i have celles taged with gfp, that is 238 amino acids long. so the probability of this happening naturally is 1/4^238 that is less than 10^-143 if you inserted dna of appropriet length. maby in a few million years this might happen, but as i saied it is a LOT faster than anyting happening naturally.

[@JK-de1oy]

Open source science is what will shape our future, thank you guys for helping speed up the process. Everyone should be allowed to create and alter anything they like if it has a positive impact on their or someone elses life.

[@typhooni3149]

Open source science is indeed the way to go.

[@mythicbiology6849]

This is a neat start to a small lab. Very much so jealous of your quick start on your DIY bio lab! I have been planning something similar for a long time, and will be looking to this upcoming series for alternative perspectives for lab processes and setups. Best of luck to you on this!

[@Assault_Butter_Knife]

Will be waiting to see you making the electrophoresis machine!

[@Artemis-zl5cs]

1:27 nice xkcd hazard symbol you got there

[@krabkit]

that comic only went up 2 days ago too

[@kitmanevo]

I am not sure if uv-c can pass-through the plastic cover of the lamp.

[@PiezowDimD]

I built a ozone generator and air ionizer powered by a neon backlight driver from a big LCD TV (old tech). It was enough to superficially sterilize during the night a room a bit smaller than yours. The air ionizer helps to trap particles on grounded surfaces. It's needles produce less ozone than the generator, allowing work while it's active.

[@roberttomsiii3728]

To anyone considering this just buy a grow tent. You can get a 4ft × 8 ft at 6.5ft tent for about $250. Most any other size can be found for just over $100 Not to say I haven't considered doing this as well but the material cost is about equal; then the time and easy of assembly as well as disassembly without harming the material investment all have value.

[@setha6096]

This is awesome! Can't wait to see how the spider silk goes.

[@mandernachluca3774]

Yeah, me to. I never heared of an amateure doing something like that. I wonder how he incorporates the DNA.

[@setha6096]

@@mandernachluca3774 He will probably use restriction enzymes to cut the desired gene out of the spider DNA. Then using the same enzymes he can insert it into the new organism's DNA. It will take some practice, and trial and error, but I think he could do it.

[@thethoughtemporium]

Nope gonna use PCR. Assuming my prep is right it should generate 9 different lengths of silk which gives me a lot more interesting stuff to work with. Restriction enzymes are way more expensive and don't work as well for isolating stuff from genomic DNA as you get cuts in weird places and you don't have as much DNA to work with in the end. Then I'll add some adapters to the ends of those fragments for insertion into a plasmid. But those will actually be cut with a restriction enzyme. I'm designing this to only need 2 restriction enzymes total. 1 for the plasmid, 1 for the gene.

[@setha6096]

Okay, that will be awesome. I can't wait to see it.

[@mandernachluca3774]

@@thethoughtemporium Very interesting, i hope you go more into detail in your upcoming video, for someone like me who has not a lot of knowledge about biochemistry it will be very hard to understand. Can't wait to increase my knowledge on that subject XD.

[@thekohhh]

What is the air filter you are using? Can I get a link to it?

[@squirlboy250]

I know this video is 4 years old but if you want cheap lap equipment check to see if a University near you has auctions. I live close to U of M "Michigan" and they have auctions all the time selling ALL kinds of great stuff from lab equipment, machinery, cars/trucks, house hold items, you name it they action it.

[@preisschild4622]

Looking forward to see your experiments!

[@TheZebinator]

Wouldn't it have been easier and cheaper to buy one of those ready to assemble grow tents? They make then pretty huge and are easy to pack up when you need to move it

[@zvpunry1971]

The UV-C lamp might damage the plastic foil. Maybe add some of the foil into a box with such a lamp and let it run for some hours/days and check it regularly to see if something changes, if the foil becomes brittle or if nothing happens at all.

[@romanbucharist4708]

This channel is Gold. You deserve a subscribe.

[@0_o913]

Hot boxing is optional

[@e0n662]

This human can do anything

[@themadhack3r431]

I mean I made a bird house back when i used to go to school. It was pretty good😐 haha. I really enjoy this channel nice to see so many other's with the same interests and hobbies.

[@HewanDemissie]

This is execellent presentation. I am planning to build plant genetic engineering laboratory in Africa, Ethiopia. This will help me for the start. Thanks.

[@roberttomsiii3728]

You should look at getting a grow tent.

[@aniksamiurrahman6365]

After this series is done, make a video on How to make a analytical chemistry lab. At least go for UV-Vis and IR spectroscope an analytical calorimeter, few separation techniques like few type of chromatography: paper, thin layer, reverse phase, HPLC etc chromatography etc.

[@thethoughtemporium]

Forgot to add spec to the list but I'm already sourcing parts for a uv vis scope and may build an ir FTIR scope at some point.

[@MakeTestBattle]

Could combine the spider silk DNA with the Kombucha bacteria to make Spider Silk Bio Leather? :D

[@thethoughtemporium]

been thinking about this a lot lately actually

[@enzov9772]

This is the one channel I’d max out on on Patreon

[@ojoemojo]

❤thank you!

[@StevenRotelli]

How is your lactose intolerance?

[@mihajlopetkovic2003]

Don't you think UV from lamp will degrade plastic?

[@bjarnivalur6330]

If the plastic is UV sensitive, then it definitely will, especially if the light is full spectrum (UV-A, UV-B, and UV-C)

[@mihajlopetkovic2003]

@@bjarnivalur6330 Yes, UV sterilizing lamps are UVC.

[@leobla744]

in his stream he said that he would replace the plastic if neede because its cheap and easy (sry for the bad english)

[@xxdustinadamsxx]

This was AMAZING.

[@fretcofretco3675]

Chop saws can usually be rotated to cut at the angle you need

[@npc6924]

1:27 Did you guys print out last Monday's xkcd? The one about radioactive high-voltage laser-emitting biohazard so that make the floor slippery

[@thethoughtemporium]

Yup! Thought it was the perfect thing for the lab and it's distributed under a creative commons license. If they ever have it in their store I'll buy a copy. Love xkcd

[@talegrinho6969]

Thanks for subtitles

[@piranha031091]

1:28 : I see someone's an avid XKCD reader! ^^ Impatiently awaiting all your future projects!

[@Gooberpatrol66]

"I recently ordered this vial of nightmare fuel" lol I'm dead

[@howardkurtus7069]

Holy shit that sounds dope, good luck with your endeavors.

[@cadentoutenhoofd9457]

Can you show how to make a bioreactor that is small

[@Praxis4RageBaiting]

I am very excited for future videos I can hardly wair

[@PEACE_BY_GENOCIDE]

i flippen love science

[@felipe69420]

Thank God please make more videos like this. I need more mad scientist how tos

[@franglish9265]

Pretty!

[@abyssstrider2547]

So you are gonna make a super armour huh? cool. Could you also make a rope the strength of a steel using spider silk? I am curious.

[@gcm4312]

I have no clue how biohacking works and will be looking forward to content on this! Super curious to learn about the process...

[@lordyellowman]

Pls continue this series

[@MrRolnicek]

As a HUGE fan of the webseries "Worm" I know that Black Widows have some great silk ... but I also know that Darwin's Bark spiders have even better. Are they too difficult to obtain?

[@thethoughtemporium]

They're too hard to get. If you can find one and send it to me I'll happily use that.

[@mlgsamantha5863]

All spider's silk is fundamentally the same, whatever minutiae make Darwin's Bark spider silk stronger than Black Widow's is going to be lost when you put the genes into yeast. There might be some species of spider who's silk genes are easier or harder to put in yeast, but that's not likely to be related to the silk's strength.

[@thethoughtemporium]

you don't know much about spider silk do you? That's about as far from the truth as possible.

[@mph8759]

Will the UVC degrade the plastic foil?

[@guilldea]

Im really hyped about this

[@hunterturk1557]

Love the videos and the progress

[@user-tv6sw3vt9q]

How would you go about adding an airlock? Data recovery services at scale aren't cheap whatsoever.

[@thefox347]

Colab with Cody's Lab, he does in-depth chemistry and would probably love to help you.

[@realityChemist]

Would be cool, but Montreal is pretty dang far from Utah

[@earthbjornnahkaimurrao9542]

realityChemist - I thought he was in Brazil?

[@realityChemist]

Well, he mention's that it's in the Montreal hacker space near the beginning, around 2:20 or so, and one of the shelves is labeled in French (la case populaire, I think it means something like "common box"). That's what I based by assumption that he was in Montreal off. In any case, Brazil is also far from Utah :p

[@jurassicdocs]

Cool

[@ishasimpson8002]

You should show more of the plastic eating fungus

[@RGplayer101]

Maybe a stupid question, but could you do open air agar work in these?

[@hullinstruments]

Jesus Christ please please please show the process to build and set up a mediocre electronics lab. So many people have shown this, but I feel you are one of the only ones with the thoroughness to really express it accurately.

[@gamereditor59ner22]

Thank you!! I need this!!!

[@GeorgeC1andonly]

Ambitious to try spider silk...

[@Dan-bq3rd]

Man I love your video, I am interested in modifying fungi DNA

[@thescienceemporium2606]

Could this lab work with bacteriophages beside the flasks etc..

[@leefless2004]

I really enjoy your videos!

[@RealHogweed]

this is so exciting!

[@jaydenspell6684]

What about insects

[@alexandercane7332]

Where’s part 2 home skillet?

[@hiih3212]

How is it over kill?

[@marzinjedi6437]

Operating rooms in America all have this design in the construction of the theaters I helped build a five room theater !

[@DoRC]

0:19 that guy looked nervous..

[@MorbidMindGamingLLC]

Can this clean room build also suffice for computer and hard drive repairs?

[@DoRC]

You may want to look at used electrophoresis machines on ebay. I picked one up to use as a hv power supply for like $50 shipped and its a pretty nice one.

[@thethoughtemporium]

Technically I already have one. I ordered the components for it before someone just gave me one, so I'm gonna make the video anyway. And all the parts only cost 20 bucks

[@DoRC]

@@thethoughtemporium gotcha. Yeah mine has been sitting on a shelf since i bought it. 5kv makes me nervous:)

[@thethoughtemporium]

.... why would you use 5kv for electrophoresis. What are you trying to pull through a gel? A whole hamster? It's supposed to run at 100v

[@MatthijsvanDuin]

Poor hamster... :_(

[@DoRC]

@@thethoughtemporium I haven't the foggiest idea. I don't know how that even works. It's a pharmacia multidrive xl. I was mistaken and it actually tops out at 3.5kv. I'd be interested for you to take a look online at it and let me know why it goes so high.

[@alexandregeraldes6734]

Where o find reagents? Name site?

[@marcusk7855]

Looks like my old grow room from back in the day.

[@ThomasAndersonbsf]

when is part II coming? or have you released it and I missed it:?

[@thethoughtemporium]

Just came out last week kzbin.info/www/bejne/sKK0Zatvp9Kegck

[@Winther83]

A tip the orb-web-weaving spiders are known to have the strongest silk in the spiderworld and some places "milk" it to study the material, if it is possible to replicate this spiders genetic "formula" if you manage to be successfull with the black widows . :) looking forward to see more of this.

[@thethoughtemporium]

orb weaver silk and black widow silk are almost identical. They rate approximately equal in strength, with the exception of the darwins bark spider since it's weirdly stronger than the rest by about 10x. But there's no sequence data for their silk and getting one of those spiders is nearly impossible

[@Winther83]

Ahh did not know that. will have to read up on those spiders as well. :) True it all depends on availability and that the sequence is there makes things "easier".

[@logismos9844]

I hope that you’re not planning on doing anything that requires sterility in here.... because sterile this room is not. Whereabouts are you located? I have a few ULPA and HEPA filters that are used in clean rooms I could lend you guys to build an iso 100 bio cabinet out of.

[@matislevesque5028]

did you say Montreal? I live in quebec.. nice to see where brewing good shit!

[@Bewleys_]

Why black widows? Can't silk be extracted from something less venomous?

[@thethoughtemporium]

Their silk is very similar to golden orb weavers and is some of the strongest in the world. I'd prefer to use darwins bark spider silk but getting one is a pain. And because black widows live in north america it's easy to get preserved specimen. I can get golden orb weaver specimens but they're dry not wet preserved, so DNA quality won't be as high. Also I found a nice sequence for black widows which makes it easy to clone out the gene I want. I was going to use golden orb weavers originally, but it was to expensive to have the gene synthesized. This way it's 20 bucks for the spiders, 10 for primers and boom I've got my gene. Vs 500 bucks to have it synthesized.

[@realityChemist]

Have you considered doing any work with M. Magneticum? (microbewiki.kenyon.edu/index.php/Magnetospirillum_magneticum) It's a microbe that takes in dissolved iron and deposits it as magnetite in its body. They seem to use it to navigate, but we can use organisms like these to grow magnetite nanoparticles of highly uniform size. They have applications in energy and oncology. Also they're just a particularly cool example of biomineralization. I guess it's less genetic engineering than general bioengineering, but they're non pathogenic and from what little I know of them I think you could probably grow them in your lab (although they seem to be a little picky about oxygen concentration).

[@KX36]

You need at least two vortex mixers so you can race them across the bench.

[@thethoughtemporium]

vortexers are weirdly expensive..... Was contemplating building one

[@benos1799]

Do have any advice for someone building a lab like this

[@Nopmord2252]

I think this is classified as a BSL-2 laboratory ?

[@k-Gonzo]

How would I get started doing in vitro pharmacological experiments? That's of more interest to me than genetic engineering tbh.

[@neyvindeleon1080]

Have you thought of extracting the bacteria that digest styrofoam from the intestines of your mealworms and creating a digestor with them?

[@thethoughtemporium]

Yup. it's on the list now that this lab is ready

[@neyvindeleon1080]

@@thethoughtemporium looking forward to it!

[@ethanmye-rs]

Idk if it's easy to edit, but you put silicone on the floor, not silicon

[@M_J_Glide_22]

you didn't have to cut your corners on angles. You could have simply made a box and still enclosed it like a house.

[@s7887177]

This make me think about breaking bad

[@zacbergart6840]

great video... I enjoyed it... but then had the thought that you are just showing how near we are to the story foundation of the game "The Division". still like the video though.

[@amanitamuscaria5863]

1:10 so do the programmers.

[@cwmd7651]

Oh man, this sounds amazing! Now I want to make one of my own and modify alge to grow super quickly! I would use it on space missions to make food and to solve world hunger! How do you get the stuff to analyze and modify DNA? It seems like that would be super expensive. Great video! Keep up the amazing work!

[@arazatliyev6564]

Live caputalism world,what you speak?

[@shahafzm]

Hey, i am very interested in all of the topics you are doing, so i have a question, what topics and degrees you have learned in your life, and where did you get all of this knowledge, becase i am planning on learning , and doing , everything you are doing, thank you, and i am looking forward for your reply :)

[@austinkunch710]

He said in a Q&A video that he started a bachelor's degree in biology but he only completed about 2 years and then quit to start what he's doing now. He said he does a lot of reading, and he has some friends in the bio and chem field. There's probably more than this but I think if you read, and can get some hands on experience somehow, you might be able to do some great work

[@stone4574]

Stupid question: if you make beer with spider silk yeast, would spidey-silk pee be a problem, or should it pass as dietarry fiber?

[@thethoughtemporium]

it's a protein so more than likely it would be like eating super chewy mushroom. You would never pee it out. It would never make it into your blood stream and certainly not through your kidneys.

[@trailblazingfive]

When can we expect a video about a practical approach to E. coli modification?

[@thethoughtemporium]

Week or two

[@trailblazingfive]

@@thethoughtemporium Thank you :3

[@AutumnPuffin]

should have gotten Darwin's bark spiders it you wanted to make artificial spider silk, would have some interesting follow-ups

[@thethoughtemporium]

Find me a pair of darwins bark spiders and pay to have it's genome sequenced and I'll happily use it. But until then I'm working with what's cheap and availiable. Also having spent many, many hours going over silk genes, the silk I'll be using is very similair to golden orb weavers and the natural silks are nearly equivalent in strength. So it'll be at least as strong as kevlar. And the mods I'll be doing to the sequence ought to boost strength further if it works the way I hope it does

[@jadirneto2130]

A room before actually entering the lab would help keeping the lab clean

[@talkingmetastasis607]

you're amazing

[@1224chrisng]

5:10 um actually, Silicon is the element, Silicone is the sealent

[@kajraske2002]

"...and getting it to run in yeast." Some strong programming vibes. Yeast Emulates Any Single Technique?

[@runforitman]

Doesn’t UVC produce O3? And isn’t O3 an asphyxiant and bad for your lungs? Are you sure that’s fine to be your only air source?

[@Basement-Science]

UVC is actually incapable of creating high concentrations of O3. There are equilibrium reactions that break down O3 under UVC light. Therefore the O3 levels in the room should be very low at all times. Basically when you can smell it it is unhealthy.

[@camilovicuna9796]

Canadian schmeckles

[@jimthegentleman2446]

Ugh why did i get an unskipabble ad that lasts 3 minutes?

[@4corander]

You shouldn't rip on hacker's carpentry work... next time, standard wood stud walls are much better, much sturdier and much easier to make. Cheers!

[@recklessroges]

Yay! The Voluntary Extinction Project would be proud, (if anyone existed after the accidental global catastrophe that comes from a lab loosely based on this one.)