Everything You Could Want To Know About PCR
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- Опубликовано: 11 ноя 2018
- PCR or polymerase chain reaction, is one of the most useful tools in a biologists tool kit. It allows you to take a DNA sample, select a part of it and make millions of copies of that section, amplifying it millions of times over. These newly grown fragments can be used as a diagnostic tool, or to generate pieces that can be used in other processes.
Today we take a deep dive into the world of PCR and explore how it works, how you design the reactions, and how they're run in the lab. In a future video we may also look at how to build the thermocycler that makes this process possible, but there are lots of great tutorials on building them already online.
We also take a step forward in the spider silk project and successfully isolate 1 of the 2 genes we'll need for the project. We're still trouble shooting the other so that will be the topic of a future video. The fragment we isolated is directly usable in our construction pathway and will be inserted to build a new new plasmid we're calling pKlac2g418. This means we're only a few more steps away from finishing our plasmid and making our first spider silks!
Resources:
Spider beer overview - • Spider Beer - Making Y...
Gel electrophoresis - www.khanacademy.org/test-prep...
Crash course chemistry - • The Nucleus: Crash Cou...
Crash course biology - • Carbon... SO SIMPLE: C...
The manga guide to molecular biology - www.amazon.ca/Manga-Guide-Mol...
Genome compiler - designer.genomecompiler.com/app
Primer3 - primer3.ut.ee
Snap gene - www.snapgene.com/
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Oh FFS! I do *not* need another expensive hobby! But how can one not fall in love with this stuff?
Dude, you are so creative in so many different fields of science. Freaking renaissance man
#renaissanceperson
Hello Justin,
I just wanted to tell you that I really love your channel, and that you've inspired me to switch from computer science to a biology major.
My entire life I KNEW I wanted to be a software engineer and I even completed a AA in IT.
But your videos (and a few others) made me realize that I have a far greater passion for plants and molecular biology.
I've enrolled at the University of Calgary majoring in plant biology, and I hope to eventually specialize in genetics.
Thank you so much for sharing your hobby and passion with the world.
but you'll need your IT skills anyways)
Dude, change the human genome, make people better, fix the world.
hows the biology degree going?
I’m not even out of high school and I’m having similar thoughts. As this comment was made 4 years ago, how did this turn out?
My simple engineer brain finally understood PCR, thank you for this explanation.
i am a biomedical engineer, not math smart to be engineer not biology smart to be biologist, just qualified enough to work at mcdonalds :D
Excellent video! Minor history-of-science quibble: Taq was isolated from a bug (Thermus aquaticus) in Yellowstone hot springs, while Pfu polymerase was later isolated from a deep-sea hydrothermal vent bug (Pyrococcus furiosus).
Oh true, my bad!
Every biologist would know the story of Taq polymerase
I love that this is meant to be easily understandable. I have looked for videos on the basics of Biotech for a while, and it seems that you have taken on that mantle for all of us. Thank you.
Edit: Have you considered a video on preventing Contamination? Thanks.
Yes. Contamination-prevention would be a fantastic piece to do!
So, "polymerase chain reaction machine" sounds cool and all, but who can we talk to about getting it renamed to "gene forge"?
Caffeinated Spider I think it would be better to use that name for a DNA synthesis machine. But the name is awesome haha
That is called a thermal cycler.
Gene forge? Who’s gene?
@@tampafreedom2237 Gene lives a few blocks down my street and has a pet iguana named Steve. He also grows watermelons in his front yard. You should visit him sometime, chill guy.
There's an old (originally Mac) game series called "Geneforge" - and you cannot make this up - produced by Spiderweb Software :D
This world needs more people like you... love what you are doing.
Nice graphics!
Also, great vidjayo, I really liked the quick pace of it
I love this video because it shows practical uses for what a majority of my BIO 101 class taught me keep making your videos they are so cool.
I've been looking for the answer on how each strand is the same length, even though extension doesn't have a stopping point beyond just cooling it down. This video gave me the answer. Thank you.
As somebody with a really messed up biology education (6 teachers in 5 years of which 2 were first-time and 1 not technically a teacher, also only until 10th grade) I really want to thank you for how understandable this video is. I was able to watch it and understand everything in one go! To be fair, I am a nerd but still: Very nice video!
I had been waiting for this. Thanks
Exelent video series. keep up the good work.
I recently decided to change my major from nursing to molecular biology and biotech research. I've always had these videos in my pocket, to watch when I'm interested. Now.. I'm super thankful for these videos. So educational, love it.
One of the best explanation in the web... Thanks
I actually do feel like i learned something from this youtube video, that explanation how to cut specific parts from dna makes perfect sense.
I love your channel guys when I see a new video always think it's worth to watch it or otherwise they don't post it
I actually understood most of that.
Belias Phyre A high schooler could understand it. Freshmen in my school take slightly more advanced biology though it’s not nearly as interesting or fun
Yeah... not so sure about that. At least measuring by the quality of students I see coming out of my local high school. They are barely literate, and some of them aren't even that. But those ones _cans play football good._
Thank you so much for that codon segment it helped me understand that a lot I was watching MIT lectures and was lost
Wonderful series!!!
I know this is late but I absolutely love how informative your videos are. I started watching this thinking I would be completely lost and lose interest. But i am glad I watched it bc I was very wrong, I always learn a lot from your channel. Thank you!
I want to get to know more of this so that I can make a lab to genetically test myself for my diagnosis of crohns possibly cure it. I also want to be able to test friends and family for diseases.
This is great !
Keep doing this videos ...
hey man, been waiting impatiently for your video.
Amazing how you learned everything by yourself nice job
Thank you so much!!!!! You have no idea how many minds youre inspiring and helping to learn with molecular techniques. Right from school kids to graduate students are benefitting from this. Thank you again, and I look forward for more molecular biology and bioengineering contents. I would also be happy to collaborate with you if you like.
You deserve far more exposure than you already get, people can really get a better understanding of Biotech from your videos, keep it up man!
Best way to do that is to share the videos and support the channel so I can keep making these videos. Glad you enjoyed!
Aaaaand now everyone knows about it because of Corona. You really went ahead 1 year with this video :)
so good !
I have been struggling to understand primer design and PCR and this finally solidified it for me!!! THANK YOU!
You're very welcome!
Cool stuff man
I'm subscribed to a lot of channels, but no channel gets me as excited as this one when I get a video notification. Unless I'm driving, I drop everything to watch new uploads. Keep up the good work!
Btw, what applications do you use the DIY heat block for instead of the PCR machine? On the surface, the PCR machine seems like a better and more controllable heat block, but I'm probably missing something
Glad you enjoy. The main reason is that the heat block fits bigger tubes for bigger reactions
@@thethoughtemporium ah, I see. What kind of bigger reactions are you going to be doing? More or less the same ones as the PCR machine?
Transformations (putting DNA into organsims), Protease digestion, gibson assembly, restriction digests, etc etc
Thanks for making this! It’s awesome seeing a intro level video that actually walks you through a complete experiment workflow.
My only complaint is that you talk so fast it is sometimes hard too keep up or take notes. I’ve used pause extensively, but a slightly slower pace verbally would be great.
Can’t wait to see more of your genetics videos!
i realy had lost the spirit to continue my studies but man this video is inspirational thx a lot keep up the good work
Good lord this is fascinating!....I can't believe it's possible to explain this online.
Yay! PCR!!!!!
So, how long until you are able to craft multiple copies of the pBlackwidowSpiderSilk?
If we're lucky, end of november. If not, december.
@@thethoughtemporium that's good I suppose. Definitely better than just water
Do a tutorial on Golden Gate Assembly
This is beautiful
i got a D in honors biology in highschool but this is so interesting. Thank you for this awesome content.
Biology has always seemed like a dark art to me. You're videos make it much more approachable. I'm excited to see where this spider silk thread goes!
Thanks for all your awesome content related to biology, definitely learning more things than from a common class
love this vid
This is so cool
A little note: Thermus Aquaticus (the taq polymerase synthetizing bacteria) was found in Yellowstone park, not in hydrotermal vents!
Love your videos very interesting and educational I have a dumb question is there a way or can make a video how to make homemade buffers from scratch or is it possible. Thank you
This is bending my brain. I am taking the shallow taxonomy/data/system analysis skill-set in life and this is getting pretty deep into bio-engibeering.
Give the next video asap im addicted :)
I'm really enjoying your videos. I hope I can give this a try one day. You're the most in-depth and accurate person I've seen in a long time. I always love seeing people help others and I get to learn some fun stuff while I'm at it. Good work.
fascinating.. reminds me of Gattaca :)
Thankyou!
Literally just finished the biotech unit in biology had to do about 30 pgs of work in a night but now I understand this vid.
Great video... I dropped out of biology and got into electronics and computing instead... It's funny to see that if I'd carried on with one I'd have been doing similar work to the other. :)
Throwback to my biotech studies (psychologist now)...this is a project I wish I could have done in uni.
Perfect
Dude I just need to tell you that you explain PCR much better than my goddamn teacher, like seriously, I was freaking out cause I didn't fully understand the process but you explained it really well
This was recommended for me. You'll have 250k views on this in a week
I like to think G and C as Gucci and A and T as at&t. Helps me remember which goes to which.
Nice video guys, it was very helpful for my project. But I couldn’t find the next video about building the empty plasmid. Someone can help me please ?
I have this as test tomorrow. What are the odds!!
*You should really make a video about how to synthesis recombinant HGH*
Great video, very helpful! My pedantic invertebrate biologist self compels me to point out that spiders aren't insects, though...
Finally The wait is over!
Ya the edit on this one took a long time. Started on Wednesday and finished this morning. I'm exhausted, but I think it was worth it
The Thought Emporium
Thank you
Can u pls provide a guide on how to perform a viral growth curve?
Ha alpha DNA! You’re in Montréal? Just FYI, their primers work fine for basic PCR but we had issues with them for other things such as sequencing. I suspect they skimp on purification steps to offer lower prices. Nice video!
Very interesting video thank you.
I'm curious tho, what's the price of the primers you bought ?
About 40-50 bucks for the whole set. That's the extra nice part about alpha DNA. They're super cheap. Sigma charges twice that.
I don't mean to be rude, but how is the research on this topic going so far? I am excited to hear more about it :)
And..... I'm picking up my first PCR machine today!
Nice
Hey Justin!
Where did u get those forceps? I'm just starting my Ph.D. in molecular work and I want to be the cool guy in the lab with some fancy forceps! Thanks and keep making videos they are inspiring and incredible educational
ebay. just look for titanium forceps, they're like 15 bucks
One thing you didn't touch on is that you can also extend your primer past the region you want it to bind to so that it will add that sequence to the end of your amplicon. This is extremely useful if you want to transcribe a region of DNA as you can amend your transcription promoter up stream of it.
I did actually briefly. There was a shot of exactly that. That'll be covered in the Gibson assembly video though as it's beyond what I could cover in one video
what happened to the spider silk project? was it put on hold/canceled, or are you still trying to find fresher spiders? I love this project and really wanna see you actually have an end product.
is there a specific place you buy buffers or any kits you could recommend
My dude you should totally stream your work on twitch for educational and financial awesomeness :p
I would be fascinated to see this live... o.o
question. if you have your gene sequence yo only need to ad the start aminoacid and the stop aminoacid to the sequence if the sequence doesnt have it and know what polymerase you are going to use
Amazing once again
I am a biotech intern, took micro, genetics, biotech 1 and 2 (certification program stuff, essentially did what TTE has been doing with some genome annotation) and cell cultures. being unpaid and only having an AS, what jobs (other than a part-time student assistant) could I qualify to continue gaining experience in the field until next fall? I'm looking for ideas, but if anything I could just be an intern again at utsoutwestern.
tl;dr I just want to know some other entry-level biotech jobs with some experience required
I work for a company that makes a small (8-well) DNA testing machine. I have no idea if they would consider hiring an intern, but it shouldn't hurt to occasionally check their website for new job listings. The company name is MatMaCorp.
@@WilliamDye-willdye thanks for the info! I searched them up and found their website, searching also had results for biotech-careers*dot*org and I found some local companies that make some interesting antibody kits. I wish the intern search functionality worked better on the site, but it has found a few great locations for a list.
thanks for the help!
This test is used as a diagnostic tool to detect emerging viral diseases. It is so sensitive that it can detect nucleic acid from dead virions.
Hi! I am not sure about what if I have my forward primer bonded to the intron side and 2 bp from the start of my exon (coding region) bonded in to the end of my fwd primer. So my question is when I start reading the codons from my exon, I start it including those base pairs included to my forward primer, or I start reading them after the primer?
Do you need to add ATP to the solution so the DNAPolymerase can copy dna?
biology is so cool
🤯 my mind is a little bit blown
Cool, I have experience with image processing. Could you please help me design a PCR machine?
Could you use CRISPR for this?
After my last semester of phage gene research, I couldn’t agree with this title more. XD
Out of the topic and all other stuff that i like to watch in your channel(and lack understanding and feel like an ape in space programme :P ), i have a GREAT Question with a HUGE Problem around the world which nails thousands of people ; `Mouldy` books. Apart from controlling moisture, apart from people stating `You know, i placed it in sunlight and all the mould is gone`, i've been yet so far trying to find a method which can `turn` old zombiefied stinky books back to live. So far, except so many of the useless ideas and rubbish spread over the net, few things i have found BACKED by academic research and paperwork; 1- Freezing to -86 Degrees (Which kills all the mould which made me win a jackpot and buy an ultra low freezer but didn't have guts to operate it yet) 2- Vacuum of Space 3- Freeze Drying and 4- Chemical Fumigation&Playing around with Toxic stuff (which is out of my reach due to dangers involved). I will chuck them to ultra low freezer (and then place them into vacuum chamber) but before i go-full-mad on all the mouldy books i have(which a library full of due to a disaster) i just want to know if you have any experiences or data you may share? Mould in books is a huge issue and not much amount of attention is given these days it seems. I've managed to find an old timer who was in the business for very long time but he was scared to chat about his techniques(thinking i might steal his job maybe) but he said he uses `vacuum of space` kinda method with other combined.I just want my 100+books back without mould and paperback until decent book scanners get a good price(or i got to DIY one!) Supporting the channel with 5 subs(shhh it is secret) with all the house equipment since you had 10k subs and great work and keep em coming!
Do you have the soul eater logo in your web browser? (Top left at 6:06 for example)
YEEESSSSSS!
Any info you are willing to share about your progress on GOES 17? Is it online yet?
It started snowing. Hard to be outside in a blizzard. So yes it's online, no I can't do anything about it unfortunately
How about creating your own IC... How hard would it be, creating integrated circuit outside of the industry?
ruclips.net/video/23fTB3hG5cA/видео.html
Didn't understand anything. But very interesting....
What is a primer dimer ?
Is this how forensic biology labs work when needing to test DNA but there's not a large enough sample? They take that tiny bit of genetic material that was recovered from a crime scene, take out the DNA, using yeast they get it to start multiplying until there's enough to test, then test it?
Or something like that?
did you try benchling?
Did you use a positive and negative control? :P
Hm could you teach yeast to produce methane from CO₂ and H₂O?
I've been trying to buy primers for a side project, but i am not associated to a lab and would like to have them delivered to my home lab. I'm wondering if you know of a company that will send me primers to a residential address? Do you have any tips?
anyone know what his thermal cycler is i’m looking to start my own lab
Minipcr.com
I think the speaker is talking too fast. This is a practical matter, so a little slower is better.
How could you use a dna data base wouldn’t every spider have different combinations of letters
finally i have a basic understanding on how this all works. thanks, now I want more !
What do you use now that Genome Compiler is gone?
benchling
@@thethoughtemporium Oh sick! That's what I got an account on. I'm brand new to this and know pretty much nothing about gene editing, but I'm watching your past Who's Gene is it Anyway streams. Do you plan to do another one of those soon?