Fast SYBR Green vs. TaqMan® Fast Advanced Master Mix (qPCR step 5)

The other videos in this series are:
Introduction to the steps of the qPCR workflow (qPCR step 1):
Quantitate RNA with the Qubit® 2.0 Fluorometer (qPCR step 3):
Superscript Vilo cDNA synthesis kit (qPCR step 4):
Amplify Sample with The StepOnePlus™ Real Time System (qPCR step 6):
Preparing the card for the Viia™ 7 with TaqMan® Assays (qPCR step 7):
Amplify sample with the Viia™ 7 Real-Time PCR system (qPCR step 8):

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Transcript

Now let's run our Taqman array card. So as you see these cards are stable at room temperature. They ship that way and you can store them that way.

And here's how the product looks when it first comes out of the box. Something of note, all the tacqman cards are bar coded with both the part number and the lot number.
I will now open up the card.

And, here is the card.I'll pop it out.

Comes with a little desiccator and you can see now this is what the taqman array card looks like. We have up here 8 different ports and each of these ports corresponds to 48 different wells. In each of these different wells is a dried down taqman assay.

The fill ports have two different holes. One is to fill, the other is an air port. So, this is a very simple procedure. You fill up each of these 8 ports. You've pipetted 8 different times and now you're going to be interrogating 384 different spots. Depending on the format of the card would depend on how many samples and how many genes you can look at. So, now we're ready to load the card and it's really quite simple.
Each of the different ports is labeled 1 thru 8 so I have the one closest to me. And the pipette-men is set to 100 micro-liters because that's the amount I'm going to put into each of the ports. And I'll also bring in my sample, which is just here.

And now I'm ready to load the card. So, what I will do is go into the larger of the fill ports. Add in the pipette tip and pipette my sample.

Finished. And now I will go into the next fill port, which is pour number two.

Add in the pipettemen. Pipette my sample. Done, it's as easy as that.

And there we go, all 8 ports are filled and we're ready to take it to the next step. The liquid now once it's centrifuged, will go down this little port and it will fill in all 48 wells for each of the different 8 ports. So, the way that we do this
Is, we bring in our centrifuge holder. So now let me take this out. I'll move the cart so you can see this. It's very easy. We say this side out.

We then place the card into here. [sound]. And then simply centrifuge it so that the liquid fills in all the different holes. Okay, now we're back from the centrifuge, and I will remove my card.

And you can see that the liquid is here towards the bottom of the fill port. The rest of the liquid, which is a sample of the master mix, is now down into the card.

And now we're ready to seal the card. I'll bring in the sealing device.

Here you can see it here. What I will do now, is add the card upside down.

There's only one way the card can fit, and now I'm going to push forward. And the card is sealed. We'll remove it from here.

Now I'm going to cut off the top. Where the fill ports were. Done, we're ready to run the card on the Viia 7."

[jscott]

IDT has the best deal right now on qPCR pre-designed assays called PrimeTime. 1/2 the cost of Taqman on a per reaction basis and updated with the latest SNP locations. Plus they have the fastest turn around time.

[Madbiochemist]

Life Tech has been getting better though, and have been supporting some nice promotions

[kirihitiana]

First time I'm seeing safety glasses when somebody is busy PCR'ing!