miRNA extraction from Cells and Tissue
miRNA Extraction
AROS has implemented Standard Operation Procedures (SOP’s) for miRNA extraction
from most biological tissues/cells, including muscle, brain, bone etc. miRNA
preparation will typically contain all RNA species smaller than approximately
70 bases. 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.
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 miRNA preparations. AROS also offers to quantify single
miRNA like e.g. MIRN24-1 or MIRN16-1 by Q-PCR. Each sample is analyzed
on the Agilent BioAnalyzer for successful extraction and all results
are returned as part of our Quality Control reporting process.
see Q-PCR
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 miRNA concentration, volume, total yield and
Agilent Bioanalyzer images. 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 miRNA quality.
Labels for Containers
All sample tubes must be unambiguously labeled with information that allow cross
reference to a sample log sheet that should be e-mailed to AROS. All tubes/containers
must be labeled using a permanent marker.
miRNA extraction from Formalin-fixed, Paraffin embedded Tissue Sections
RNA Extraction
AROS has implemented Standard Operation Procedures (SOP’s) for
miRNA extraction from Formalin-fixed, Paraffin embedded Tissue Sections.
Reagents employed at AROS for this is High Pure RNA Paraffin Kit from
Roche.