Pinpoint™ Slide RNA Isolation System II

Allows for the isolation of total RNA from fresh and/or FFPE tissue sections.
Simple procedure combines Pinpoint™ tissue sampling technology with a one-step RNA extraction/purification method.
Omits the use of organic denaturants.

Product Size Catalog # Price Qty
Pinpoint™ Slide RNA Isolation System II 50 Preps. R1007
$268.00

About Pinpoint™ Slide RNA Isolation System II

The Pinpoint™ Slide RNA Isolation System II is an innovative product for the isolation of RNA from any targeted area of fresh or paraffin-embedded tissue sectioned onto a glass slide. The system combines powerful Pinpoint™ tissue sampling methodology, a unique single-step RNA extraction/binding buffer, and Fast-Spin column purification technology to yield high quality RNA. Unlike current UV-based methods, this product makes isolation of tissue RNA simple and quick. No expensive specialized equipment is needed. Eluted RNA is well suited for subsequent RNA analyses including RT-PCR.


Format Spin Column.
Processing Time 5 hr
Equipment Microcentrifuge
Sample Sources Cells from tissue sections on glass slides.
RNA Recovery Typically, up to 5 µg RNA is eluted into ≥ 8 µl RNase/DNase-free water allowing for a highly concentrated sample.
RNA Storage Recommended that 1 U/10 µl RNase inhibitor be added to the RNA prior to storage at -70°C.

RT-PCR of RNA recovered from human tissue using the Pinpoint™ RNA Isolation System. Amplicons (in duplicate) are from A) a human ß-actin transcript; B) an arbitrary human transcript from Chromosome 3. M is 100 bp DNA Marker (Zymo Research).

I.   Paraffin Removal from the Tissue Sample

  1.  Mount the paraffin-embedded tissue section (≥10 µm thick) onto a glass slide and dry it at 60°C for 30 minutes.

 2.  Submerge the slide in xylene at room temperature for 1 hour changing the xylene once after 30 minutes.

 3.  Hydrate the sample by washing progressively for 2 minutes in 100%, 70%, 50% ethanol, and then pure water.

 4.  Air-dry the sample on the slide.  RNA isolation using the Pinpoint™ Slide RNA Isolation System II can now be performed.

 

II.  Pinpoint Fractionation

            (Procedure for the removal of a selected area of tissue from a glass slide.)

 1.  Apply the Pinpoint™ Solution1 to the area of tissue on the slide where the RNA is to be extracted2.

Use a sterile pipette tip or a syringe to gently spread a small amount of Pinpoint™ Solution over the selected tissue region.  Generally, use about 0.5 µl of Pinpoint™ Solution per mm2 of tissue area

 2.  Allow the Pinpoint™ Solution to dry completely at room temperature. (Usually about 30 to 45 minutes).

The PinpointTM Solution should dry as a blue film embedding the tissue and cells underneath.

3.  Remove the embedded tissue from the slide.

Use a sterile blade or scalpel to cut, and then remove the embedded section from the slide.  Transfer the sample to a 1.5 ml tube.

4.  Centrifuge briefly to locate the tissue sample at the bottom of the tube.

III.  RNA Extraction

(Procedure for the extraction and purification of total RNA from a deparaffinized tissue sample.)

 1.  Add 20 µl of RNA Digestion Buffer and 5 µl Proteinase K to the tube containing the recovered tissue.  Mix gently.

For multiple samples, the RNA Digestion Buffer and Proteinase K may be premixed. Add 25 µl of this mixture to each sample.

2.  Incubate the tubes at 55°C for 4 hours.

3.  Centrifuge the tubes briefly when the incubation is finished.

4.  Add 50 µl (2 volumes) of RNA Extraction Buffer and mix.

5.  Add 75 µl (1 volume) of 95-100% ethanol to the tube.  Lightly vortex.

6.  Transfer the mixture to the Zymo-Spin™ IC Column in a Collection Tube.

7.  Spin the column at ≥10,000 x g for 1 minute.

8.  Add 200 µl RNA Wash Buffer to the Zymo-Spin™ IC Column and centrifuge at ≥10,000 x g for 1minute.  Discard flow-through.  Repeat this step.

9.  Transfer the column into a new RNase-Free Tube.

10.  Add 10 µl of prewarmed DNase/RNase-Free Water (60°C) directly to the column matrix.  Wait for 2 minutes then centrifuge at ≥10,000 x g for 1 minute and collect the eluted RNA.  The RNA can be used immediately or stored at -70 oC.

For specific notes and additional information, please see the product protocol PDF.
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