Video 2 v2 - PCR and Cloning

Aug 08, 2026 08:06 · 4:21 · English · Whisper Turbo · 2 Ομιλητές

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S… Speaker 1 (Video 2 v2 - PCR and Cloning)

Hello and welcome back to our quest to prove that we can make a vaccine for canine

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S… Speaker 1 (Video 2 v2 - PCR and Cloning)

coronavirus in just 100 days.

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S… Speaker 1 (Video 2 v2 - PCR and Cloning)

This week,

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S… Speaker 1 (Video 2 v2 - PCR and Cloning)

the viral genes being used to make the vaccine arrived a whole day ahead of schedule,

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S… Speaker 1 (Video 2 v2 - PCR and Cloning)

so the team was in a flurry to get everything ready for the next steps.

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So basically this week we received the gene box we ordered and all

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S… Speaker 2 (Video 2 v2 - PCR and Cloning)

our primers.

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S… Speaker 2 (Video 2 v2 - PCR and Cloning)

So we've been able to start our planning process.

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S… Speaker 2 (Video 2 v2 - PCR and Cloning)

So getting the inserts ready for all the different constructs

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S… Speaker 2 (Video 2 v2 - PCR and Cloning)

that were designed.

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S… Speaker 2 (Video 2 v2 - PCR and Cloning)

So we started that yesterday.

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S… Speaker 1 (Video 2 v2 - PCR and Cloning)

So how does this tiny amount of raw genes get processed not only into

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S… Speaker 1 (Video 2 v2 - PCR and Cloning)

a vaccine, but into enough vaccine for hundreds of dosages?

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S… Speaker 1 (Video 2 v2 - PCR and Cloning)

It all starts with gene cloning.

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S… Speaker 1 (Video 2 v2 - PCR and Cloning)

You may have heard of cloned animals like Dolly the Sheep or seen movies and

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S… Speaker 1 (Video 2 v2 - PCR and Cloning)

TV shows where they're cloning genes.

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S… Speaker 1 (Video 2 v2 - PCR and Cloning)

But the real process into making a vaccine is a completely different story.

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S… Speaker 1 (Video 2 v2 - PCR and Cloning)

The first step is to take these gene blocks and rehydrate them.

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S… Speaker 2 (Video 2 v2 - PCR and Cloning)

So we're cloning basically the fusion protein,

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S… Speaker 2 (Video 2 v2 - PCR and Cloning)

so the specific small part of the virus that we

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S… Speaker 2 (Video 2 v2 - PCR and Cloning)

use to...

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S… Speaker 2 (Video 2 v2 - PCR and Cloning)

pre -act our antigen,

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S… Speaker 2 (Video 2 v2 - PCR and Cloning)

so it's not the whole virus,

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S… Speaker 2 (Video 2 v2 - PCR and Cloning)

it is a subunit of it,

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S… Speaker 2 (Video 2 v2 - PCR and Cloning)

and it is basically that sequence combined with the molecular

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S… Speaker 1 (Video 2 v2 - PCR and Cloning)

clamp.

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S… Speaker 1 (Video 2 v2 - PCR and Cloning)

Now,

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here comes the cool part,

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S… Speaker 1 (Video 2 v2 - PCR and Cloning)

taking our gene construct and amplifying it.

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S… Speaker 1 (Video 2 v2 - PCR and Cloning)

If we wanted to make,

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S… Speaker 1 (Video 2 v2 - PCR and Cloning)

say, thousands of copies of a mug,

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S… Speaker 1 (Video 2 v2 - PCR and Cloning)

we would have to have a factory to make them.

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S… Speaker 1 (Video 2 v2 - PCR and Cloning)

But when we're making copies of our genes,

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S… Speaker 1 (Video 2 v2 - PCR and Cloning)

we use E.

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S… Speaker 1 (Video 2 v2 - PCR and Cloning)

coli as our cellular machines.

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S… Speaker 1 (Video 2 v2 - PCR and Cloning)

Bacteria like E.

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S… Speaker 1 (Video 2 v2 - PCR and Cloning)

coli have the ability to uptake genetic material in the form of plasmids,

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S… Speaker 1 (Video 2 v2 - PCR and Cloning)

copy it for their own use,

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S… Speaker 1 (Video 2 v2 - PCR and Cloning)

and spread it to other E.

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S… Speaker 1 (Video 2 v2 - PCR and Cloning)

coli.

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S… Speaker 1 (Video 2 v2 - PCR and Cloning)

It's like the plasmid is a DVD with an awesome movie on it,

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S… Speaker 1 (Video 2 v2 - PCR and Cloning)

and all of the E.

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S… Speaker 1 (Video 2 v2 - PCR and Cloning)

coli are pirating that movie and then passing it along to their friends,

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S… Speaker 1 (Video 2 v2 - PCR and Cloning)

creating thousands and thousands of copies of our movie in the process.

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S… Speaker 1 (Video 2 v2 - PCR and Cloning)

So all we need to do is make our genetic material in the form of a

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S… Speaker 1 (Video 2 v2 - PCR and Cloning)

lot of DVDs for.

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S… Speaker 1 (Video 2 v2 - PCR and Cloning)

To make enough plasmids for our movie to go viral or maybe bacterial,

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S… Speaker 1 (Video 2 v2 - PCR and Cloning)

we need to use PCR or a polymerase chain reaction.

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S… Speaker 2 (Video 2 v2 - PCR and Cloning)

PCR is an incredibly big part of what we do.

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S… Speaker 2 (Video 2 v2 - PCR and Cloning)

So whenever we create an antigen,

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S… Speaker 2 (Video 2 v2 - PCR and Cloning)

which is the small part of the virus that we use that we put in

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S… Speaker 2 (Video 2 v2 - PCR and Cloning)

the molecular clamp.

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S… Speaker 2 (Video 2 v2 - PCR and Cloning)

Sometimes you need to do slight modifications for it to make it more stable

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S… Speaker 2 (Video 2 v2 - PCR and Cloning)

or to make sure it expresses better.

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S… Speaker 2 (Video 2 v2 - PCR and Cloning)

So we have our gene blocks,

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S… Speaker 2 (Video 2 v2 - PCR and Cloning)

which are basically almost a native sequence,

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S… Speaker 2 (Video 2 v2 - PCR and Cloning)

and then we use different primers to change small things in it.

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S… Speaker 2 (Video 2 v2 - PCR and Cloning)

And that stands for a PCR.

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S… Speaker 1 (Video 2 v2 - PCR and Cloning)

Usually when cells are dividing,

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S… Speaker 1 (Video 2 v2 - PCR and Cloning)

they need to copy their DNA first with the enzyme polymerase.

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S… Speaker 1 (Video 2 v2 - PCR and Cloning)

What this reaction does is take a polymerase from a bacteria native to hot

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S… Speaker 1 (Video 2 v2 - PCR and Cloning)

springs and put it in a solution with our gene block that we need to copy.

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S… Speaker 1 (Video 2 v2 - PCR and Cloning)

We add in primers,

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S… Speaker 1 (Video 2 v2 - PCR and Cloning)

small segments of DNA that matches where we want polymerase to

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replicate and acts as a homing beacon for the polymerase.

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S… Speaker 1 (Video 2 v2 - PCR and Cloning)

Because the polymerase is from a hot spring bacteria,

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S… Speaker 1 (Video 2 v2 - PCR and Cloning)

we can warm up the DNA to split it for replicating,

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S… Speaker 1 (Video 2 v2 - PCR and Cloning)

cool it down so the primers can attach,

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S… Speaker 1 (Video 2 v2 - PCR and Cloning)

and then warm it back up again so that the bacterial polymerase can duplicate it all.

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S… Speaker 1 (Video 2 v2 - PCR and Cloning)

Each cycle of this doubles the DNA,

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S… Speaker 1 (Video 2 v2 - PCR and Cloning)

so we quickly get enough DNA for all of our plasmids.

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S… Speaker 1 (Video 2 v2 - PCR and Cloning)

Then, after we check the PCR copied the correct sequences,

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S… Speaker 1 (Video 2 v2 - PCR and Cloning)

we cut and glue the different sections together into a plasmid using a bunch

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S… Speaker 1 (Video 2 v2 - PCR and Cloning)

of other enzymes.

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S… Speaker 1 (Video 2 v2 - PCR and Cloning)

It's like we're editing the movie,

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S… Speaker 1 (Video 2 v2 - PCR and Cloning)

cutting and ordering the scenes to make a movie that we can put onto our DVD.

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S… Speaker 1 (Video 2 v2 - PCR and Cloning)

Then we get our DVDs,

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S… Speaker 1 (Video 2 v2 - PCR and Cloning)

our plasmids,

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S… Speaker 1 (Video 2 v2 - PCR and Cloning)

and we put them in a solution with our E.

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S… Speaker 1 (Video 2 v2 - PCR and Cloning)

coli, heating the solution so that the plasmids are uptaken.

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S… Speaker 1 (Video 2 v2 - PCR and Cloning)

Now, all that was left was for the team to let all the E.

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S… Speaker 1 (Video 2 v2 - PCR and Cloning)

coli grow on agar plates and incubate.

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S… Speaker 1 (Video 2 v2 - PCR and Cloning)

And every time they replicate,

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S… Speaker 1 (Video 2 v2 - PCR and Cloning)

they're copying our plasmid and sharing it.

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S… Speaker 2 (Video 2 v2 - PCR and Cloning)

Then go through the process of transformation where we put

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S… Speaker 2 (Video 2 v2 - PCR and Cloning)

the DNA into E.

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S… Speaker 2 (Video 2 v2 - PCR and Cloning)

coli cells.

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S… Speaker 1 (Video 2 v2 - PCR and Cloning)

It's through this process that the team turns a tiny little pellet of

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S… Speaker 1 (Video 2 v2 - PCR and Cloning)

DNA into enough genetic material that next week...

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S… Speaker 1 (Video 2 v2 - PCR and Cloning)

they can start turning these genes into protein for the vaccine.

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S… Speaker 1 (Video 2 v2 - PCR and Cloning)

At the moment,

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S… Speaker 1 (Video 2 v2 - PCR and Cloning)

we're currently transforming those into E.

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S… Speaker 1 (Video 2 v2 - PCR and Cloning)

coli bacteria,

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S… Speaker 1 (Video 2 v2 - PCR and Cloning)

which we all played on agar plates overnight and purify out the DNA.

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S… Speaker 1 (Video 2 v2 - PCR and Cloning)

Then we will transfect their DNA into trocells.

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S… Speaker 1 (Video 2 v2 - PCR and Cloning)

But sadly,

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S… Speaker 1 (Video 2 v2 - PCR and Cloning)

we have to wait a while for all of that E.

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S… Speaker 1 (Video 2 v2 - PCR and Cloning)

coli to grow.

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S… Speaker 1 (Video 2 v2 - PCR and Cloning)

So that's it for today.

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S… Speaker 1 (Video 2 v2 - PCR and Cloning)

Tune in next week to see what the team does with all of that E.

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S… Speaker 1 (Video 2 v2 - PCR and Cloning)

coli on their journey to create a vaccine in less than 100 days.

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S… Speaker 1 (Video 2 v2 - PCR and Cloning)

See ya!

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