General Information
Total RNA isolation services are available for cultured cells, blood and harvested tissue. Cells and tissue are disrupted and homogenized in TRIzol® Reagent (Invitrogen) or QIAzol™ Lysis Reagent (Qiagen), nucleic acids retrieved by Chloroform extraction, and total RNA purified using the Qiagen EZ1® Advanced. Tissues that require more vigorous physical manipulation are homogenized using the Qiagen TissueLyser II®.
Total RNA isolation from blood is performed using the PAXgene™ Blood RNA System (PreAnalytiX). This method requires specific sample collection tubes available from PreAnalytiX and specific handling instructions.
Sample requirements
All samples submitted for RNA isolation should be prepared in the appropriate vessel (see below), properly labeled with a legible sample identifier. An AGTC project submission form and a full sample list prepared in MS Excel must be emailed to Daniel Lott prior to sample submission. All samples must be stored at -80°C and transported to the AGTC on dry ice.
Cells must be removed from culture and submitted to the AGTC in nuclease-free, 1.5 or 2.0 milliliter (ml) microcentrifuge tubes. Samples prepared in lysis solution (TRIzol® or QIAzol®), should follow the product specifications for volume per number of cells, to a maximum of 1 ml per sample. Alternatively, cells can be prepared in RNAlater™ RNA Stabilization Reagent (Qiagen).
Tissue submitted for RNA isolation can be flash frozen or preserved in RNAlater™ RNA Stabilization Reagent. Flash frozen samples should be approximately 0.5 cm x 0.5 cm x 0.5 cm in size and submitted in nuclease-free 1.5 or 2.0 ml microcentrifuge tubes, frozen on dry ice. Tissue preserved in RNAlater™ RNA Stabilization Reagent should be approximately 0.5 cm x 0.5 cm x 0.5 cm in size and submitted in nuclease-free vessel appropriate to accommodate the required volume of preservative (please refer to the RNAlater™ Handbook), frozen on dry ice.
Whole blood submitted for RNA isolation must be collected in PAXgene™ Blood collection tubes in accordance with the PAXgene™ Blood RNA Isolation protocol.
Additional Information
If needed, please contact Dan Lott for further information regarding RNA isolation.