Isolate RNA with the PureLink™ RNA Mini Kit (qPCR step 2)





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Published on Apr 12, 2011

The PureLink™ RNA Mini Kit provides rapid purification of total RNA from a wide range of cell and tissue types, including animal, plant, and yeast cells, bacteria, and blood. PureLink™ technology combines guanidine-isothiocyanate lysis with the speed, purity, and ease of use of silica-membrane purification. The safe and easy procedure can be completed in less than 20 minutes without the need for hazardous phenol/chloroform extraction, CsCl centrifugation, or LiCl or alcohol precipitation. High-quality purified RNA can be obtained from mini- to midi-prep amounts of starting material, with minimal genomic DNA contamination. Purified RNA is ready to be used in downstream applications such as RT-PCR, qRT-PCR, northern blots, cDNA synthesis, and nuclease protection assays.

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


Okay. Let's unbox our QPCR work flow starting with preparing the sample.
The kit we'll be using today is called the Pure Link RNA mini kit. Pure Link RNA mini kit is a fantastic kit to purify RNA from cells or tissues. It's a very easy-to-use kit and a very quick kit. So, essentially it just requires pipetting and centrifugation steps and in 20 minutes you will have isolated your RNA. Let's see how it works.

We'll open this up. And take out the quick reference card.
Now, in here, we have some tubes, we have the lysus buffer.Wash buffer one.Wash buffer 2.

And finally, RNAse free water. In addition to the chemicals, we also have the recovery tubse.The collection tubes.
And finally the spin cartridges. The way that this work flow works is that we will first of all, we will take our sample, which has been lysed and homogenized and we add it to a collection tub. Allow me to show that. We will pipette into the tap. And now the RNA will bind to the spin column. We will centrifuge and wash this several times.

I happen to have a tube here that's already been centrifuged. So, once we've added the cell lysate to the collection tube, the RNA will bind to the spin column. And we will centrifuge such that the liquid comes down into the collection tube, as you see here.

Now after the 3rd wash we are going to take the top of the tube off, and insert it into a collection tube. So now we're going to add water to the top and alute the RNA with water into the bottom of this collection tube.

We take it to the centrifuge...centrifuge it. And then, here's the final product. We have water in the bottom. With the RNA, we will remove the spin column, close this, label it, and we are done with our RNA isolation.

For more information, please visit: http://products.invitrogen.com/ivgn/p...


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