AquaPreserveTMis
an aqueous solution-based preservative with DNA, RNA, and protein
extraction functionality. It may be used to streamline blood sample
collection, stabilization, transport, storage, and DNA, RNA, and
protein extraction. The extraction protocols are simple. There is no
need to remove the plasma or lyse the RBC or fractionate the blood
cells prior to extraction. Cell-free DNA, RNA, and proteins will not be
lost and gene expression profiles will not be altered artificially by
in vitro separation of blood components and cells. Most uniquely,
AquaPreserve can be used to extract intact total RNA from existing
frozen blood samples collected in regular anticoagulants, thus enabling
the use of surplus and archived frozen blood samples collected during
routine medical care and clinical trials for blood RNA research.
Blood is the most collected biospecimen, however,
there is no RNA kit for extracting intact RNA from
archived frozen whole blood collected in regular anticoagulants.
With AquaPreserve, you can extract total RNA from
fresh or frozen whole blood: (1) lyse whole blood with
AquaPreserve, (2) pellet hemoglobin with ProSink, and then (3)
precipitate DNA and RNA with isopropanol.
Collect blood in EDTA
vacutainers and store them at -80°C.
Use no RNA
preservative-containing PAXgene or Tempus tubes.
Extract
RNA from population or disease-targeted blood samples for
prospective studies.
Extract
RNA from surplus or archived blood samples for
retrospective studies.
Recover not
only cellular but also cell-free DNA and RNA in whole blood.
Require
no WBC fractionation or RBC lysis or protease
digestion of hemoglobin.
Extract DNA, RNA, and
proteins from both fresh and frozen whole blood.
Use no additional
and different DNA, RNA, and protein extraction kits.
Preservation
and extraction of blood DNA and RNA with AquaPreserve.
Frozen human ACD blood was thawed in 2 vol
of AquaPreserve. Aliquots of the AquaPreserve lysed blood were stored
at -80, 22, and 37°C for 7 days. DNA and RNA were recovered
from the cleared lysate by isopropanol precipitation. One tenth of the
total DNA/RNA from 200 ul of whole blood was digested with DNase I and
electrophoresized in a 0.8% non-denaturing agarose gel.
Features
Compatible with conventional blood draws
AquaPreserve is compatible with anticoagulated
blood samples collected in routine medical care and clinical trials. The blood
samples can be used for other clinical tests and only an aliquot of the extra
or leftover blood samples are saved in AquaPreserve for DNA and RNA extraction.
Safeguard the blood samples during long-term storage
AquaPreserve-stabilized blood sample can be stored long-term at -80 °C without
risking RNA degradation due to accidental blood thawing during storage, such as power outage or
freezer
breakdown. Unlike plain frozen blood, AquaPreserve-stabilized blood
sample can
be repeatedly thawed to give out aliquots for distribution or experiments, a significant advantage and advancement in
blood biobanking.
Extracting
DNA, RNA, and proteins without using other extraction kits
AquaPreserve not only stabilizes but also
extracts DNA, RNA, and proteins without using additional extraction kits. It streamlines blood
collection, transport, storage, and DNA/RNA/protein extraction, reducing pre-analytical
variation and processing cost.
Comparison
Comparison
of Techniques for Biospecimen Preservation
Formalin
R*later
AquaPreserve
Mechanism of Stabilization
Cross-linking
High
salt dehydration
Enzyme
inactivation
Add
to Frozen Specimens
No
No. Require pre-soaking
Yes
Storage
Temperature
22
°C
22 °C
22 to
-80 °C
Damage
to Specimen
Yes
No
No
Inactivation
of Pathogens
Yes
No
Yes
Offensive
Odor
Yes
No
No
DNA,
RNA, and protein Extraction
No
No
Yes
Gross
anatomy of liver tissues preserved in AquaPreserve and PBS. Fresh mouse
liver tissues (~100 mg) were submerged in 0.2 ml AquaPreserve (left) or
PBS (right) and stored at 22 °C for 7 days. As seen, liver
tissues preserved in AquaPreserve looked fresh, remained intact, and
wedged in the tube, while liver tissues stored in PBS became highly
degraded and sank to the bottom of the tube.
Brief Protocol
1. Lyse the blood cells
Add
0.25 ml of AquaPreserve to 0.25 ml of fresh or frozen whole blood in a
1.5-ml microfuge tube. Vortex or shake to mix well or until the frozen
blood sample is completely thawed in AquaPreserve (Do
not thaw the frozen blood without mixing with AquaPreserve or the RNA
will be degraded during blood thawing. However, if you are only
interested in blood DNA extraction, the blood sample may be thawed,
aliquoted and then mixed with AquaPreserve.).
2. Remove the proteins
Add
0.125 ml of ProSink (#9030) to the AquaPreserve lysed blood. Vortex for
1 min to mix well and incubate at 22 °C for >30 min (Blood DNA
is now stable at 4-22 °C for a year, and blood RNA is now stable at
-80 °C forever, 4 °C for months, 22 °C for 2 weeks, and 37 °C for 2 days.).
Centrifuge at 3,500-14,000 xg for 10 min to pellet the proteins.
3. Pellet the DNA/RNA
Pour the supernatant (~0.8 ml) to a
1.5-ml microfuge tube preloaded with 0.8 vol (~0.64 ml) of isopropanol
and vortex to mix. Centrifuge at 3,500-14,000 xg for 10 min to pellet the
DNA/RNA. Decant to discard the supernatant and rinse the DNA/RNA pellet
with 70% ethanol twice. Suspend the DNA/RNA pellet in 50 ul of TE buffer or deionized
water.
Use the volume ratio of 1:1:0.5 (blood:AquaPreserve:ProSink) for other extraction scales
Micro
Mini
Midi
Maxi
Blood (ul)
50
250
2,000
4,000
AquaPreserve (ul)
50
250
2,000
4,000
ProSink (ul)
25
125
1,000
2,000
Centrifuge tubes
0.6-ml
1.5-ml
15-ml
15-ml
DNA yield (ug)
2-3
12-15
100-130
200-250
RNA yield (ng)
50*
250
2,000
4,000
Extractions per #8060 and #9030
1,200
240
30
15
Price per extraction ($)
0.21
1.03
8.27
16.53
* This is the RNA yield from human blood. For mouse blood, RNA yield may be 2.5 ug/50 ul blood.
Ordering Information
Buy-1-Get-1-Free Promotional Sales!
We will add a free unit of every AquaPreserve and ProSink ordered as a thank-you to our customers.
Product
Name: AquaPreserve Kit
Product Number:8001, 8060
Application:Biospecimen preservation and
extraction. For in vitro research use only.
Size:
The kit is sufficient for the preparation of
8001: 4 DNA/RNA
minipreps (use
0.25 ml AquaPreserve for 0.25 ml blood)
8060: 240 DNA/RNA
minipreps
Kit
Contents: The AquaPreserve Kit
includes the following items
8001: 1 ml
AquaPreserve Solution, Instruction Manual
8060: 60 ml
AquaPreserve Solution, Instruction Manual
Price:
8001: $10 each
8060: $199 each
Ordering:
To order, please
click the "BUY" button below (order ProSink #9030 separately).
