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.
|Processing Time||1.5 hr|
|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.
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 Pinpoint™ 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 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.)
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.