Blood extraction
Isolation of DNA, RNA
and microRNA
Our services cover the extraction of DNA and
RNA from blood using Standard Operation Procedures (SOP’s) based
on manual methods or robotic extraction. If needed, DNA, RNA
and miRNA can be extracted from the same blood sample. All
extractions are in a quality recommended for many downstream
processes including microarrays and Q-PCR. All steps are performed
under GLP compliant conditions and if needed under formalized GLP.
DNA extraction from whole blood
RNA extraction from whole blood
miRNA extraction from whole blood
DNA extraction from whole blood
Blood Collection for DNA isolation
AROS isolates DNA from fresh or frozen whole blood (with common
anticoagulants such as citrate and EDTA) using an IVD approved
Robot with a fast automated 96-well–plate procedure. Using this
technology, genomic and mitochondrial DNA of high quality are
achieved with sufficient yields and ready to use in downstream
applications like PCR, blotting procedures and SNP microarray
analysis. see
technical report
Storage of Blood Samples
If the blood samples need to be stored before shipping to AROS
they should be frozen at -20ºC for short term storage, and at -80ºC
for long term storage immediately after blood drawing.
DNA Extraction
AROS has implemented Standard Operation Procedures (SOP’s) for
DNA extraction from whole blood on the MDx Biorobot from Qiagen Inc.
Quantitation / Qualitation
DNA yield is determined on a Beckman Coulter DTX880 Platereader
or a Nanodrop (ND-1000 Spectrophotometer) by measuring optical density
at A260 nm wavelength. The A260/A280 nm wavelength ratio is accessed
and should be 1.8 – 2.0 for high quality DNA preparations. Each
sample is checked using the Agarose gel chromatography and all results
are returned as part of our Quality Control reporting process.
DNA Yield
Typical yield of DNA is 4-10µg per 200µl of whole blood,
which is sufficient for microarray analysis using AROS SOP’s in
which 1µg DNA is needed for the labeling reaction. Blood DNA for
microarray.
Blood Tube Labels
All tubes must be labeled unambiguously using a permanent marker.
Reporting
The Investigator is provided with a printout and electronic file
including the A260/A280 nm wavelength ratio, the DNA concentration, volume,
total yield and agarose gel image. The customers’ own standard
report scheme may be provided to AROS for reporting.
Sample storage
Following extraction and purification, DNA is stored at -80°C. AROS
offers to bank the samples within AROS for a period of 6 months
free of charge. For long term storage see
Sample Banking
Notification
A sample log sheet must be received with all samples. A standard
sample log sheet can be e-mailed to AROS to be used as report
template; otherwise AROS’ standard
report sheet will be used
RNA extraction from whole blood
RNA Extraction
AROS has implemented Standard Operation Procedures (SOP’s) for RNA extraction
from blood on the MDx Biorobot from Qiagen Inc.
Quantitation / Qualitation
RNA yield is determined on a Beckman Coulter DTX880 Platereader 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.
RNA Yield
AROS guarantee a minimum yield of 3µg of RNA per PAXtube (in >95% of
the samples) as long as the tubes has been correctly filled with 2.5 ml of whole
blood, and subsequently mixed thoroughly. This yield is sufficient for microarray
analysis using AROS SOP’s in which 1µg RNA is needed for the labeling
reaction. Blood RNA for microarray
Blood Tube Labels
Preferentially, all sample tubes should be labeled with Barcodes (vertically
on the middle of the tube) for optimal sample tracking during processing. If
no barcodes are available all tubes must be labeled unambiguously using a permanent
marker.
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 and 28S/18S ribosomal RNA ratio from the Bioanalyzer. The customers’ own
standard report scheme may be provided to AROS for reporting.
Sample storage
Following extraction and purification, RNA and DNA is stored at -80°C. AROS
offers to bank the samples within AROS for a period of 6 months free of charge.
For long term storage see Sample Banking.
In order to ensure RNA quality we recommend that you follow some basic procedures
for blood collection and storage using the PAXgene technology. Please inquire.
Notification
A sample log sheet must be received with all samples. A standard sample log sheet
can be e-mailed to AROS to be used as report template; otherwise AROS’ standard
report sheet will be used.
miRNA extraction from whole blood
miRNA Extraction
AROS has implemented Standard Operation Procedures (SOP’s) for
miRNA extraction from blood on the MDx Biorobot from Qiagen Inc.
Quantitation / Qualitation
RNA quantity is determined on a Beckman Coulter DTX880 Platereader
or 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.
Blood Tube Labels
Preferentially, all sample tubes should be labeled with Barcodes
(vertically on the middle of the tube) for optimal sample tracking during
processing. If no barcodes are available all tubes must be labeled unambiguously
using a permanent marker.
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 Bioanalyzer image. The customers’ own
standard report scheme may be provided to AROS for reporting.
Sample storage
Following extraction and purification, miRNA is stored at -80°C.
AROS offers to bank the samples within AROS for a period of 6 months
free of charge. For long term storage see Sample Banking.