Pinpoint™ Slide RNA Isolation System I

Allows for the isolation of total RNA from fresh 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 I 50 Preps. R1003
$162.00

About Pinpoint™ Slide RNA Isolation System I

The Pinpoint™ Slide RNA Isolation System I is an innovative product for the isolation of RNA from any targeted area of fresh 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 1.5 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.  Preparation of Tissue Sections

1.  Use a sterile ethanol/water solution to clean glass sample slides and then dry by autoclaving or baking at 300°C for 4 hours.
2.  Mount a tissue section (≥10 µm thick) onto a glass slide and dry it at 60°C for 30 minutes.
3.  Submerge the slide in 95% ethanol at room temperature for 60 minutes to fix the section.
4.  Air dry the sample on the slide for approximately 30 minutes.  RNA isolation can now be performed using the Pinpoint™ Slide RNA Isolation System I.

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 the Pinpoint™ Solution over the selected   tissue region.  Generally, use about 0.5 µl of the 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 PinpointSolution 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 then remove the embedded section from the slide.  Transfer the sample to an RNase-free tube.

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

 

III.  RNA Extraction

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

  1. Add 200 µl of RNA Extraction Buffer to the tube containing the embedded tissue sample.
  2. Lyse the embedded tissue sample by pipetting the RNA Extraction Buffer up and down.  Vortex briefly.
  3. Incubate the sample on ice for 30 minutes vortexing briefly every 10 minutes.
  4. Add 200 µl ethanol (100%) to the sample, mix and then incubate on ice for 10 minutes.
  5. Transfer the mixture to the the Zymo-Spin™ IC Column in a Collection Tube.
  6. Centrifuge column at ≥10,000 x g in a microcentrifuge for 1 minute.  Discard the flow-through.
  7. Add 200 µl RNA Wash Buffer to the column and centrifuge at ≥10,000 x g for 1 minute.  Discard the flow-through.  Repeat wash step.
  8. Transfer the column to a new RNase-free tube.

Add 10 µl DNase/RNase-Free Water1 directly to the column matrix.  Wait for 1 minute 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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