RNA extraction

RNA extraction from Cells and Tissue

RNA Extraction
AROS has implemented Standard Operation Procedures (SOP’s) for RNA extraction from most biological tissues/cells, including muscle, brain, bone etc. Reagents employed at AROS are mainly TRIzol reagents from Invitrogen, and column based technologies from Qiagen Inc. Tissue specimens are homogenized in Qiagen Tissuelyzer homogenizer. AROS recommend its own in-house SOP’s for RNA isolation, or use methods specifically requested by the customer.

RNA Purification
On request from the customer an on-column DNase I digestion will be performed on previously extracted RNA. This procedure is performed in order to remove residual genomic DNA where this is critical to downstream processes (e.g. SAGE, Differential Display, some Q-PCR applications). RNeasy purification may also be performed on previously extracted RNA in order to remove residual proteins where this is critical to downstream processes.

Quantitation / Quality Analysis
RNA quantity is determined on a Nanodrop (ND-1000 Spectrophotometer) by measuring optical density at A260 nm wavelength. The A260/A280 nm wavelength ratio is accessed and should be 1.9 – 2.1 for high quality RNA preparations. Each sample is confirmed for integrity using the Agilent BioAnalyzer and all results are returned as part of our Quality Control reporting process.

Sample storage
Following extraction and purification, the RNA is stored at -80°C. AROS offers to bank the samples within AROS for a period of 6 months free of charge, or for an extended period see Sample Banking

Reporting
The Investigator is provided with a printout and electronic file including the A260/A280 nm wavelength ratio, the RNA concentration, volume, total yield, and RIN, as well as 28S/18S ribosomal RNA ratio from the Bioanalyzer. The customers own standard report scheme may be provided to AROS for reporting.

Tissue
In order to maximize the probability of success, we recommend that you follow some basic procedures to ensure RNA quality.

Labels for Containers
“All sample tubes must be unambiguously labeled with information that allows cross reference to a sample log sheet that should be e-mailed to AROS. For labeling we recommend using barcodes or a permanent marker.

RNA extraction from Formalin-fixed, Paraffin embedded Tissue Sections

RNA Extraction
AROS has implemented Standard Operation Procedures (SOP’s) for RNA extraction from Formalin-fixed, Paraffin embedded Tissue Sections. Reagents employed at AROS for this is High Pure RNA Paraffin Kit from Roche. The yield of RNA is typically 1-1½ µg per cm2 of a 10µ tissue section. The main fraction of isolated RNA is in the size range of 150-250 bases.