Extracting Spider/Bacteria DNA Using Columns – Spider Silk Step 1

Extracting Spider/Bacteria DNA Using Columns – Spider Silk Step 1


This video is sponsored by
The Great Courses Plus Doing biology work is a lot like cooking You’re always following a recipe
but there are countless potential tweaks and variations you can make
to change the final outcome With biology every technique
has half a dozen variations And the ingredients are made by
dozens of manufacturers each with their own custom blends
of compounds for the task at hand Think of biological reagents like
barbecue sauce A recipe may just call for barbecue sauce
but there are dozens of brands and flavors and your choice can play
a big role in how the final dish tastes So, most lab work requires some knowledge of which protocol variations and particular products work for your specific experiment Take electrophoresis for example Using the same basic recipe you can perform
all of these different blotting experiments on a huge assortment of molecules or with PCR there’s likely
dozens of different types based on the DNA source and
what you’re trying to do to it As well as a ton of other subtle factors What usually ends up happening
is you shop around look at all the different
products and protocols and see if you can find one that
says it does the thing you’re trying to do Then you order those supplies,
try it out, and see what happens. Maybe it works right off the bat but often you’ll end up having to
try a couple others before you find one that gives you good results Today we’re going to look at
one central recipe And some of the variations we’ve
been using over the past few months If you’ve been following along,
you’ll know that we’re working on engineering a strain of yeast
to produce spider silk One of the first steps in that process is
getting some DNA out of the spiders so we can isolate
the silk gene itself This has proven to be
a nightmare and to date, I’ve used almost a dozen spiders
in various attempts So today we’re going to explore and compare
successful DNA extraction procedures for two different DNA sources Some bacteria And the spiders Before we go further let’s go over the basic
cake recipe before we talk about flavours we’ll be using a column based approach which tends to give better results than the shot glass
and alcohol method you may have seen before It’s not to say that
that can’t work but this method is more gentle and keeps the DNA intact better as well as being
standard lab procedure there are also other variations,
like bead-based methods and we’ll look at some of those
in future videos All of these extractions work
on a similar series of steps First, we get the thing we want to
extract DNA from and make it into a sort of paste
and add a salty, soapy solution to it this pops open all the cells so that the
DNA is floating around in the salty water we then remove as much of the debris as we can be it bits of broken cells,
or a leg, or whatever else This is done using a centrifuge, and all the
heavy debris will sink to the bottom We then take our clean solution
and load it into this special tube It’s basically the world’s
smallest coffee filter and the filter, in this case, is either
made of silica or a special resin we load the tube into a larger tube which will act as our collection cup and then spin it in the centrifuge to
pull the liquid through the filter DNA is negatively charged and in
the salty conditions of the solution the silica or resin becomes positively charged so the two stick together when we spin down the column the rest of the solution containing everything
else just passes right through we then do a series of washes
to clean everything and then we’re ready to collect the DNA we transfer the filter to a clean tube and then add a small amount
of either sterile, distilled water or various other solutions so long as they’re not salty without the salt, the silica’s charges
get hidden and the DNA falls off this time when we spin everything down we end up with a tiny amount of
concentrated DNA solution most of the time when you do
this you buy all the tubes as a kit that come with all the different
solutions you’ll need the manufacturers tune the solutions in
each kit to work for their particular application so if you try to mix and match,
your results can be pretty poor right now I have two different kits and I even tried mixing up my own solutions
though that didn’t work very well Let’s start with the bacterial kit This one is made to isolate plasmids which are small circles of DNA that we
add to bacteria to make them do new things sort of like putting a CD into a computer in this case I grew some E. coli that have a
plasmid in them that makes them bioluminesce but this should work regardless
of what the exact plasmid is the plasmid uses tetracycline
as the selection agent so I prepared 1 milliliter of LB broth with 10 micrograms per milliliter of tetracycline and innoculated it with some bacteria
and let it grow overnight on the day of the extraction I first spun down this liquid
to collect all the bacteria into a little pellet at the bottom the LB was removed and then it was on to the
various buffers and solutions in the kit The kit just labels things like
“PD1” “PD2” and “W1” which doesn’t really tell you what’s going on but I was able to find some more info online PD1 contains an enzyme called RNAse which, as the name implies,
destroys RNA this is to make sure you only isolate DNA PD2 is what’s called a Lysis buffer basically, it’s very salty, and contains a
special soap or surfactant in this case SDS as well as an enzyme, called Lysozyme which eats bacterial cell walls the salt makes the bactera shrink,
the enzyme eats away at the structual integrity of the bacteria and since cells are basically living bubbles of oil the soap makes them all pop open and whatever was contained inside is released into the solution you add both of these to your bacterial pellet and then re-suspend the bacteria
and let it sit for a few minutes so it has some time to work Once it’s done we add PD3 Which seems to crash out all the
proteins and debris from the solution making it go cloudy We spin this down, which collects all the floating
junk at the bottom, leaving our DNA in solution then we carefully pipette
off the upper liquid called the supernatant and run it through the filter using the centrifuge After it’s been run through the column
we wash the filter with wash buffer which contains a large amount of alcohol
to help remove any stubborn organics we then spin the filter while
it’s empty for a few minutes to dry it so all the ethanol is gone
before we elute the DNA For the elution, we add 50 microliters of
sterile water or elution buffer since the kit comes with that Rather than spinning it down immediately, we wait five to 10 minutes to give
the DNA time to release from the filter I’ve left columns for half an hour or more
if I really want the DNA to come off Then we spin it down to collect our sample You can actually run a second
batch of elution buffer through or collect the first batch and run
it through again to try and squeeze out
every last bit of DNA But that’s it, and the extraction is complete It’s pretty slow because of all the
waiting for things to spin but it can be done in less than an hour
once you get a rhythm going As I mentioned earlier, there are some
other variants of this same basic protocol and some forego the columns and instead you magnetic beads coated
in either silica or the resin Replacing some of the centrifuge steps with beads
can really speed this up That’s why I was working
on making some of my own I actually just got the supplies to make more So expect some tests of those soon Now, let’s compare to the spiders I actually tried running a spider through the bacteria kit and it failed as spectacularly as you might imagine The reason is because the solutions
are tuned to solve a different set of problems First, lysozyme, the enzyme in the bacteria kit
only affects components of bacterial cell walls So the much more robust spider cell walls
and clumpy proteins were unaffected On top of that, the spiders are full of
natural inhibitors and other enzymes that are still active So when you pop open any of the cells they can wreak havoc and destroy any
good DNA you manage to get out So I picked up a special insect/arthropod kit which addresses all of this It’s a 27 step procedure and some of these steps loop
or have long incubation times When I did this I started at 5pm
and didn’t finish until 11:30 not time efficient by any means
but if it works, it works First up, it only wanted 30 milligrams of tissue Which is about a third of a spider This may not seem like a lot, but
it’ll give plenty of DNA to work with I chose part of the dorsal side of the abdomen As it’s high enough enough to not touch the silk glands and make a mess but also contains lots of soft tissue this was suspended in CTL buffer and I used a P1000 tip to grind the tissue
and disperse it in the solution in this kit, this is the lysis buffer but instead of SDS as their soap
they use a cationic surfactant called CTAB Then I added an enzyme called Proteinase K As a protease, its job is to break down proteins And since spiders are chock full of them
it’s got its work cut out for it To speed things along the tube was put into a heat block set to 60 degrees Celsius This is the longest part of this process and can take anywhere from 30 minutes to 4 hours Before putting it in, the solution was murky and opaque but after two hours it had fully cleared which is the stopping point To really clean things up, the kit says to add a 24:1 mixture of chloroform and isoamyl alcohol Neither of which come in the kit So I made some chloroform and sourced the isoamyl What this will do is collect any non-polar debris as well as twist proteins inside out and move them out of the aqueous layer After giving everything a gentile mix The tube is spun down and
you end up with a layer of debris at the interface between
the water and chloroform I ended up making a stack of stuff so I could get this at eye level to make it easier Not great lab practice but I wanted good results The top aqueous layer is transferred to a fresh, sterile 1.5 milliliter tube for further processing Being careful not to disturb the debris To our now clean solution we add
some RNAse A and HBC buffer and incubate at 70 degrees Celsius for 10 minutes This will destroy any RNA while also deactivating any proteins that stuck around then we add ethanol before loading into the same sort of filter and tube setup as before However in this case the kit says their
filters are resin-based not silica based which I thought was interesting After spinning that down, it’s on to a series
of washes with the provided wash buffer And finally after all that we can dry the column by spinning it empty for a few minutes Transfer it to a final collection
tube and elute our DNA Again, letting it rehydrate for a few minutes beforehand As you can see it’s still the same basic process
but because of the tough tissue sample extra steps and ingredients were
needed to get the best results Speaking of the results, there are two ways to analyze your sample once the extraction is done Run it on a gel, which we’ve
discussed in two previous videos Or test it with a special spectrometer Personally, I like using a gel better Not only is it vastly cheaper, since we dye the DNA with a stain that really only sticks to DNA It’s much harder to get a false positive
which actually happened during an earlier spider extract The spectrometer said we had hundreds of nanograms of DNA But when we ran it on a gel,
there was clearly no DNA present This is because the spectrometer can easily be set off by protein or other chemical contaminants left over from the extraction We’ve since run both samples on a gel and we finally got lots of DNA out of the bacteria and spiders We actually saw the bacterial
result in the gel dock video But there’s a problem with the spiders In theory we’re supposed to be isolating genomic DNA which is huge strands of DNA As we discussed in the gel electrophoresis video this should look like a nice
band near the top of the gel But the spider DNA looks like a smear low on the gel, which means its highly damaged and most of what we isolated is
shredded DNA fragments This actually matches up with an earlier result where we managed to get a whiff of spider DNA but at the time assumed it had just been damaged by the extraction protocol Now the running hypothesis is that the ethanol preserved the spiders and prevents them from rotting But it doesn’t stop the spiders native
enzymes from destroying the DNA And this kind of makes sense When I cut into the spiders they
were mush on the inside meaning everything was damaged So now I’m looking to source fresher spiders. I’ve already got a few leads, so
hopefully some of them will turn out As a last resort, we may switch to another spider species because they’re easier to get But for now I want to see if we
can get the widows to work At least now we know that this kit works so as soon as we have a fresher specimen we can definitely move forward but other than that, those are all the basics of DNA extraction using columns Depending on the style and exact kit the amount of time it takes can vary, but
it’s never really a difficult process It mostly involves planning in advance, getting the right tools, and following the recipe Before we wrap up this is actually a great moment to talk about the
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NileRed I know some of you already watch his videos but he did a great video on extracting
DNA with the shot glass method Though he used a whole one liter beaker. So be sure to check that out if you want to see how you can extract DNA without using any fancy kits and using only stuff you have
lying around the house And that’s where I think I’ll end this video If you enjoyed be sure to subscribe and
ring the bell to see when I post new videos. While sponsors for videos are great, this channel is made possible largely because of my patrons and channel members. So if you’d like to help the research and keep the videos coming, consider supporting. That’s all for now!
And I’ll see you next week!

100 Comments

  • luckylion

    January 11, 2019

    10:15 I think the spider is probably not well conserved, thus the degraded DNA. It might be that the ethanol did not get into the spider well enough due to the strong cuticular. I worked with lot's of insects for DNA extraction, and as long as they are conserved in 96% ethanol (and the ethanol is exchanged with fresh 96% after a few hours) the specimens give good DNA. The replacement of ethanol can be important as the specimens contain water that dilutes the ethanol. A ethanol concentration of 70-80% might lead to DNA degradation over time. Not sure here, since I would think most enzymes would be denatured by the ethanol, so maybe it's just "normal" tissue / DNA degradation? Not sure here what exactly drives DNA degradation, but 96% ethanol should not lead to DNA degradation. Great video as always = )

    Reply
  • Nothing

    January 11, 2019

    Warm dark places tend to have arachnids.

    Reply
  • Mask 1O1

    January 11, 2019

    0:59 is that a iBook G4?

    Reply
  • Kevin L

    January 11, 2019

    Too bad you are not in the USA… We have some invasive Latrodectus geometricus that are incredibly abundant and would be easy to source from Southern California with zero qualms about reducing native spider populations. If you know what specifics would need to be satisfied on the legal front that may be a viable option for you to look into…

    Reply
  • MuX1 Sam

    January 11, 2019

    like before watching

    Reply
  • jerry coleman

    January 11, 2019

    How many live ones do you need ?

    Reply
  • Repaired

    January 12, 2019

    Happy to see you got a sponsor

    Reply
  • Siana Gearz

    January 12, 2019

    I'm sorry to say this, you're the only channel that i have this sentiment towards, because most people speak way too slowly for me, but: could you please, please try to speak slower by approximately 10%. I know there are playback speed controls, but speeding up always works fine, while slowing down makes a really garbled mess that is unintelligible to me. And keep in mind, if i didn't like what you're doing or what you have to say, i wouldn't be complaining.

    Reply
  • ItsJTS

    January 12, 2019

    when are you doing another super worm Styrofoam update video

    Reply
  • Kadar Laszlo

    January 12, 2019

    Is there any chance to make a fiber like rayon

    Reply
  • Kyle Gervers

    January 12, 2019

    Would it be possible to lyophilize frozen spiders prior to extraction? I work with semi-recalcitrant plant tissues (conifer needles and twigs) and I've found that it makes specimens wonderfully brittle, allowing for more thorough homogenization.

    Reply
  • Kyle Gervers

    January 12, 2019

    You can also store your spiders for a fairly long time in CTAB. This is often what biologists do while collecting specimens in the field.

    Reply
  • X Builder

    January 12, 2019

    Heck, I have HUNDREDS of rather large black widows (and several other spiders of the Theridiidae family) just hanging round my house and woodpile. Shall I box a few of em up on ice? lol

    Reply
  • Jesse C

    January 12, 2019

    We strive to understand microorganisms and we once were microorganisms, kind of interesting.

    Reply
  • Isaac Miles-Watt

    January 13, 2019

    Quote out of context: “about a third of a spider”

    Reply
  • Ddelop Kwapn

    January 13, 2019

    WIsh you would have done a quick segment on making chloroform in the middle of the video so I won't have it on my search history lol.

    Reply
  • Collin Smith

    January 14, 2019

    Don’t ever feel bad about using black widow spiders for DNA samples. Use as many as you need (and then some).

    Reply
  • hikiwi

    January 14, 2019

    Yaaay! Sponsor!!!

    Reply
  • deus vult

    January 15, 2019

    duuuuuuuuuudededed duuuuuuuddeedede
    ddddddddddddddddddddddddddddddddddddddddddddddddddddddddddddddddddddddddddddddddddddddddddddddddduuuuuuuuuuuuuuuuuuuuuuuuuuuuuuuuuuuuuuuuuuuuuuuuuuuuuuuuuuuuuuuuuuuuuuuuuuuuuuuuuuuuuuuuuuuuuuuuuuuuuuuuuuuuudddddddddddddddddddddddddddddddddddddddddeeeeeeeeeeeeeeeeeee!!!!!!!!!!!!!!!!!!

    Reply
  • TruePowerIsMine

    January 16, 2019

    Will you consider being a guest on my show, I find your research fascinating.

    Reply
  • cringe boi

    January 16, 2019

    Watch out for peta

    Reply
  • a_wandering_silhouette

    January 18, 2019

    your nanodrop readout at 9:28 has a horrible 260/280 value. you'd expect this ratio to be 1,8-2 and 0,56 already tells you that there is wayyy more protein remaining than normal. not saying that i wouldnt have looked past that red flag and still used the DNA in hopes of a successful PCR though. also, i dont think you necessarily need one of these kits to isolate the DNA. ive had success extracting DNA from eucaryotic cells by simply boiling them for 30 mins with some proteinase K thrown on top and you can isolate DNA from mouse tail tips for genotyping with a very very simple protocol, no kit involved. maybe you could try a simple protocol such as this https://www.ncbi.nlm.nih.gov/pubmed/20204529 (bypass evil springer with sci-hub) and try adding different proteinase K concentrations to the buffer since you already have it. dont forget to kill it afterwards though, you dont want to throw your polymerase in there. also, maybe try freezing the spider in lN2 and crash it with a hammer – doesnt get any more fine than that.

    Reply
  • Prasetyo Muhammad Dwi

    January 18, 2019

    you can get the purity by A260/280 in spectrophotometer

    Reply
  • Aravind Karthigeyan

    January 19, 2019

    How are you going to isolate the silk protein gene from the spider?

    Reply
  • ElxCriiO

    January 20, 2019

    Your channel need more subs, amazing content

    Reply
  • Trae Taylor

    January 21, 2019

    can someone explain to me why exactly you would want to make spider silk?

    Reply
  • MrKittke

    January 21, 2019

    I study biology at university and this is way more intresting than most things we see!!!
    Also i think you could use a jack , same jacks they use in chemistry , they are quite inexpensive even 3d printable , and then you no longer need to stack stuff to work on eye level 😉
    Also couldn't you get live spiders from like a local spider keeper?

    Reply
  • realityChemist

    January 21, 2019

    Sorry your extraction hasn't been working. Still, when the main problem you're having is trouble sourcing fresh enough black widow spiders, you know what you're working on is really cool.

    Reply
  • Ratchet4647

    January 22, 2019

    Funnily enough.
    This semester my BioLab is working on a project where we catch spiders, extract their DNA, and sequence it in order to create phylogenetic trees.
    The day before yesterday we caught like 50 spiders from a few species at a local park.
    No black widows but tons of some more common species.
    Unless there's something different to black widow spider silk, I recommend you find out what the most common species is in your area and go out and catch a few.
    That way the DNA isn't all digested by the time of the extraction.

    Reply
  • jake lee

    January 23, 2019

    maybe go catch your own black widow knock them out with co2 and then do what you need to do.

    Reply
  • Bismuth LD

    January 24, 2019

    Spider breeding.

    Reply
  • Ismael Goldsteck

    January 28, 2019

    Is this your main job?

    Reply
  • Crypto Nein

    January 30, 2019

    What do you think about this guy's kits and such: http://www.the-odin.com/ ?

    Reply
  • Maracachucho

    February 8, 2019

    This may be well beyond the capabilities of your lab, and I could be wrong, but wouldn't it be better if you splice the spider's silk genes into the common silk worm instead of into yeast? Theoretically at least.

    Reply
  • Pasdor 993

    February 10, 2019

    How do you make chloroform, I’m asking for a friend

    Reply
  • Gavin Kelson

    February 14, 2019

    Why are these videos out of order 😂 but they're amazing

    Reply
  • Illia Kukharenko

    February 17, 2019

    Dude, it's just so cool!

    Reply
  • krizerlene

    February 20, 2019

    There is alot of smart people here

    Reply
  • Tony Sanders

    February 27, 2019

    Goats have been GMO to make silk from their milk

    Reply
  • Tony Sanders

    February 27, 2019

    Goats have been GMO to make silk from their milk

    Reply
  • Savage Strike

    February 27, 2019

    Is there a gene that gives the spiders their scopulae en how can you do a extraction on them

    Reply
  • floridatechgrl

    February 28, 2019

    The plasmid kit shouldn't work for genomic DNA of any kind. When you "crash" out the proteins with the third buffer. The genomic DNA should be in tact enough that the genomic DNA itself will "crash out". Since the plasmid DNA is smaller it will stay in solution. So just heads up that that kit shouldn't work for yeast or bacterial genomic DNA.

    Reply
  • Kadar Laszlo

    March 10, 2019

    you isnt make more part?IM WAITING

    Reply
  • naminogiri

    March 14, 2019

    Look at this
    https://youtu.be/Ds8ZFzOwGeI
    Why refine the spider silk protein instead to use it directly in a material generated by the bacteria?

    The experiment made by this guys in converting bacteria colonies in synthetic skin could benefit greatly of your work

    Reply
  • Autoboti T.

    March 25, 2019

    I am so glad I found your stuff!! So cool!

    Reply
  • kiril6656

    April 4, 2019

    Hi, In this method at what rmp do you spin the columns?

    Reply
  • AffordYourNuance

    April 6, 2019

    Great video, I am learning a lot. Couldn't you just order an oligo of the spider gene for silk and use that in your experiment with the yeast?

    Reply
  • kai ohitsuji

    April 28, 2019

    For What you will use this dna?

    Reply
  • GoldenFox

    April 29, 2019

    Why widows and now orb weavers because pound for pound orb weavers spin much more silk and there bigger

    Reply
  • Genki G

    April 29, 2019

    I still have my first gel I made during my studies, me and my partner where the only ones to get it right, might have altered the steps a bit which the teachers didnt like too much.

    So I dropped out.

    Reply
  • brett Frank

    April 30, 2019

    Is there a way that I can get spider DNA other than by hand

    Reply
  • minethree58

    April 30, 2019

    Might be a bit late, but I live in northern texas and there are black widows EVERYWHERE here. If you want to get in touch I could ship a couple live ones over.

    Reply
  • Jai Carlson

    May 1, 2019

    We are insanely lucky that this person is a nice mad scientist.

    Reply
  • S. J.

    May 5, 2019

    'Cells are basically living bubbles of oil.' Can I write that in my exam?

    Reply
  • Machinofacture

    May 6, 2019

    what are you planning to do after extracting the DNA? if it's PCR, then it might still work because chances are you have TRACE amounts of longer DNA that contain the full length gene.

    Reply
  • Чё Бля

    May 7, 2019

    > science is like cooking
    More like "making a witch brew in a cauldron".
    In that case, brew from spiders.

    Reply
  • Alexandru Ivan

    May 7, 2019

    Just a possible idea. Due to Spiders having specialized cells that create this spider silk, those cells create the silk in high quantities. Your Ecoli is not optimized for spider silk production. You should try and simplify your plasmid or use a high copy number plasmid, as well as extracting 2-3 spiders at a time.

    Reply
  • Robert Reynolds

    May 9, 2019

    I know where a bunch of black widows are, I know at least two places you can go and catch all your heart desires! Are you anywhere near Arroyo Grande, California (San Luis Obispo County)? I can take you to the shed when I'm next down that way, BUT I'M NOT GOING IN WITH YOU! OMG I HATE SPIDERS! And I know it's in my mind, because I'm more afraid of big ugly spiders that aren't dangerous than I am of Black Widows and the Brown Recluse. (I just realized, my ex was sometimes a brown recluse when he was with me and if I had died he would have become a black widow(er)!
    HAHAHA!

    Reply
  • Tanner Myers

    May 12, 2019

    I was a cook for 4 years while in college, now im going to grad school for biochemistry 😮

    Reply
  • Parker Shields

    May 13, 2019

    Hey, I work in an arachnology lab. If you are wanting just a couple of southern widows (L. mactans), you are welcome to the couple of live ones I keep. If you need larger quantities (like 100 or more), I'll have to collect them closer to the fall when they are more plentiful.

    Reply
  • Graham Fox

    May 14, 2019

    If you want some black widows from Florida PM me

    Reply
  • Nicoló Cappagli

    May 20, 2019

    I'm looking forward to see the next video.

    Reply
  • Rose

    May 21, 2019

    w hat

    Reply
  • Dailydosesofgrass

    May 22, 2019

    7:53 you just made some cloroform? There has to be a law against that… Right? RIGHT?!

    Reply
  • Curtis Cole

    May 28, 2019

    Your gonna be a real Spider-Man first geckskin and now webs the only thing left is a harness system and the shooters

    Reply
  • shafthespaceegg

    May 29, 2019

    Why are you extracting genomic DNA from the spiders instead of extracting RNA and reverse transcribing into cDNA? Does the gene encoding spider silk have introns? If so will the yeast be able to properly splice the resulting transcript properly once you ligate the spider silk gene into a plasmid and transform it into the yeast?

    Reply
  • Jackson Herriott

    May 30, 2019

    I cringed when you pipetted more than 100µl straight onto the column membrane not the wall…….

    Reply
  • Tarantula LP

    June 4, 2019

    If I put the CBD/THC genes into yeast and brew beer with it, or bake something, would I get high?
    Also, would these fungi be illegal in a country where cannabis is illegal?

    Reply
  • Aaron Payne

    June 13, 2019

    All of your videos are great, and this one is no different… But spiders and cake in the same video, your killing me man. What did cake ever do to you.

    Reply
  • Marian

    June 16, 2019

    I am extracting DNA of a weevil and I get as shmear in my gel too. HMM I will look into getting fresher samples

    Reply
  • Hermod Sass

    June 17, 2019

    Can u make a guide to genetic manipulation

    Reply
  • Nic Tanghe

    June 18, 2019

    Part 2 ?

    Reply
  • Pete

    June 18, 2019

    did you use a refrigerated centrifuge? Im guessing no. most plasmid purification procedures say spin the columns at 4 Celsius

    Reply
  • Unlimitless Power

    June 28, 2019

    So I want to extract dna from my baby brother should I cut off a toe and make a paste from that or grind up the whole thing

    Reply
  • sarah

    July 7, 2019

    imagine what would be possible if biologists had infinite stuff to make things with

    Reply
  • Rafael Esteves

    July 8, 2019

    Can you explain to me why DNA is negatively charged? It makes sense to me that if DNA is an acid (Desoxiribonucleic Acid) it should be positively charged…

    Reply
  • Gazz Maz

    July 17, 2019

    Will it blend
    Plus also putting random body parts in one tube will contaminate the dna so use the body parts you actually need
    Mixing eye dna in a hybridization project can destroy your results

    Now ill get back to making stupid comments

    Reply
  • Kineticwizzy

    July 17, 2019

    I literally have no clue what just happened

    Reply
  • William Shreckengost

    July 17, 2019

    I learned basic programming skills and how to build robots of questionable utility entirely because of the Web, and gained a hobbyist's competency in 3D modelling and CNC macguffin building/operation almost solely thanks to YouTube and Reddit posts. I still find it just plain amazing to see you confidently talking about genetic engineering and doing the work. You bet I clicked the bell.

    Reply
  • Joseph Alvin

    July 20, 2019

    To really preserve DNA in samples include a chelator like EDTA. DNAses almost always require Mg to work, RNAses are more difficult.

    Reply
  • Dr. Paul Plumbing Heating

    July 20, 2019

    Your as interesting brain wise as I am,,,, what about a crisper? Amino acids to proteins I think is 16 or 17 Amino acids,, your good,, I'm rusty practice on this stuff because I owned a plumbing heating business locally that would get me home by 6pm for dinner,, glad to study with you..

    Reply
  • Max Turner

    July 28, 2019

    You and nile red are my favorite youtubers <3

    Reply
  • Brandon Nehiley

    July 31, 2019

    Will their be anymore videos on this subject since I can't find any way of putting them in proper order and never got to see if it ever worked

    Reply
  • Mike Rogers

    August 7, 2019

    Really like your videos man they're excellent! 2 questions: 1 – I would love to build a small lab myself but it's prohibitively expensive for me… just a specrophotometer alone is a couple grand at least. Do you own all that equipment, and which suppliers would you recommend are good value for what they produce? 2 – What do you intend on doing with the spider silk if you manage to mass produce it using yeast? I finished my undergrad in biochem 4 years ago but have since moved over completely to tech, so would love to revisit biology/chem perhaps as a hobby on the side once funds allow for it

    Reply
  • Didier Codere-Lortie

    August 12, 2019

    Met his dad he is my new neighbours. After talking for 4 minutes he goes: "my kid is a genius and doe youtube for a living, I'm like yeah sure"… then this happened….

    Reply
  • rixogtr

    August 18, 2019

    Mate, do you even sleep sometimes ?
    😀
    Awesome content, I don't think I know a youtuber that is working on so many projects from so many categories 😀

    Reply
  • Mr. Black

    August 31, 2019

    If i were to do this, id use a species i could raise in captivity, and use captive bred individuals for all manor of research. Behaviors, anatomy, genetics, ect. By genetics i mean stuff like this, but also heritaty and what not. Possibly other kinda of research into them but thats besides the original point, which was that id use a captive population as to not withdraw from the wild stock and have more control over the dna i use.

    Reply
  • TheOrganicartist

    October 8, 2019

    If you want to capture the DNA without letting enzymes get a chance to damage it look into obtaining some liquid Nitrogen and cyro freeze the live spider before harvesting the tissue section you process for DNA.

    Reply
  • Sergeant Shultz

    November 5, 2019

    are you a software engineer, an electrical engineer, or a biologist?

    Reply
  • Inspector Steve

    November 15, 2019

    How did you make Chloroform? Also are you doing this out of your home or at work?

    Reply
  • John Davis

    November 16, 2019

    Is he going to become spiderman next!?

    Reply
  • Electra Flarefire

    November 22, 2019

    *looks at the dozens of golden orbs in the Australian back yard here* Do you want some? Or is this project over?

    Reply
  • Grammar Guru

    November 24, 2019

    Clean your dna if you get weird nanodrop results…. bruh

    Reply
  • bob mike

    December 6, 2019

    “…so i made some chloroform…” you know, just casually

    Reply
  • Silent Parrot Studio

    December 8, 2019

    So… if we inject that we become like Spiderman? 😀

    Reply
  • Paulie's Channel

    December 10, 2019

    Wow you'll be cloning people soon 👀👀👀

    Reply
  • Martin Verrisin

    December 12, 2019

    he often says "DNA circles" … that confuses me, I thought DNA is a single string… yet he makes it seem like there are tons of different DNA circles next to each other?

    Reply
  • Martin Verrisin

    December 12, 2019

    Is black widow silk really the best? If not, why use those? – Just use any spider. It seems crazy to me someone would buy spiders, with the amp supply everybody already has…

    Reply
  • jally fajardo

    December 13, 2019

    this is cool i am a sientist of DNA thank you for give ing me examples

    Reply
  • Rachel Madison

    December 28, 2019

    Hi, has there been an update on this project? If you're still looking to procure spiders would redback spiders work? There's tons of people in Australia who deal with infestations of them, leokimvideo for example. I'm sure if you reached out on social media you could find someone willing to trap and ship you some fresh spiders. Right now it's the summer over there, perfect time to round up some fresh ones.

    Reply

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