Amplify Sample with The StepOnePlus™ Real Time System (qPCR step 6)

The StepOnePlus™ Real-Time PCR System is a 96-well Real-Time PCR instrument perfect for both first-time and experienced users. The StepOnePlus™ Real-Time PCR System can be setup in a variety of configurations and comes ready to use, out of the box, with intuitive data analysis and instrument control software. Utilizing robust LED based 4-color optical recording, the StepOnePlus™ Real-Time PCR System is designed to deliver precise, quantitative Real-Time PCR results for a variety of genomic research applications.

The other videos in this series are:
Introduction to the steps of the qPCR workflow (qPCR step 1):
   • Introduction to the steps of the qPCR...  
Isolate RNA with the PureLink™ RNA Mini Kit (qPCR step 2):    • Isolate RNA with the PureLink™ RNA Mi...  
Quantitate RNA with the Qubit® 2.0 Fluorometer (qPCR step 3):
   • Quantitate RNA with the Qubit® 2.0 Fl...  
Superscript Vilo cDNA synthesis kit (qPCR step 4):
   • Superscript Vilo cDNA synthesis kit (...  
Fast SYBR Green vs. TaqMan® Fast Advanced Master Mix (qPCR step 5):
   • Fast SYBR Green vs. TaqMan® Fast Adva...  
Preparing the card for the Viia™ 7 with TaqMan® Assays (qPCR step 7):
   • Preparing the card for the Viia™ 7 wi...  
Amplify sample with the Viia™ 7 Real-Time PCR system (qPCR step 8):
   • Amplify sample with the Viia™ 7 Real-...  

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Transcript:

Alright. Let's recap a typical real-time PCR or qPCR experiment in a plate. What you will require is the actual plate, the plastics and the plastic holder.

The optical adhesive cover and the optical adhesive sealer.
In addition for the reagents, if you're doing a Taqman experiment, you will require the Taqman fast advance master mix.

TaqMan gene expression assay.

Sybr experiments will require the sybr ring master mix, and, 2 primers, which I have designated, primer 1 and primer 2. P 1 and P 2.

Also. When we fire your CDNA sample and finally some water.

These pipe [indiscernible] are to show you that you essentially pipette the reagents into the plate. You then seal the plate. This is the final product, which we then run on the instrument and in this case the Step One Plus. And now I'm walking over to the Step One Plus. I'm going to load my plate in the instrument simply by pulling open the door.

And setting the plate, The A goes up in the top as usual.
And closing the door. Now I'm just going to use the touch screen function of the Step One Plus. And I already have TaqMan Fast CDNA protocol right here on the main menu. So, I am just going to choose that one and press save.
OK I saved my experiment. I start the run now.

Say okay. Now what's happening is, the cover is heating. And as soon as the cover has come up to the proper temperature, you will see that this wide band becomes smaller as the plate is moving up. And as it says here on the screen, the required run temperature of the heated lid is 105. We're right now at 90.5. So after a few more degrees, the run will begin. So now that the run is completed on the step 1 plus, I'm going to collect results by putting in my USB drive, and hitting the collect results button. The system is busy, it's writing the USB. Please don't remove the USB device.

And you can see as the data is being written, we have the little bars coming across the screen. Now my experiment is ready. I may remove my USB storage. I hit okay. My data is on here. I'm going to go take it to my computer at my desk and analyze my results.

For more information, visit: http://www.invitrogen.com/site/us/en/...