AquaPreserveTM is a multifunctional reagent for DNA/RNA/protein preservation and extraction. It may be used to streamline biospecimen collection, stabilization, transport, storage, distribution, and DNA/RNA/protein extraction. By streamlining the entire biospecimen workflow, AquaPreserve can reduce pre-analytical variability, prevent cross-contamination, and increase data reproducibility and reliability. Furthermore, AquaPreserve extracts total DNA/RNA/proteins/metabolites from the whole blood; it recovers both cellular and cell-free circulating DNA/RNA/proteins/metabolites in the whole blood, therefore, maximizing the scientific value and utilities of the blood biospecimens.
Streamline biospecimen pre-analytical workflow
AquaPreserve not only preserves but also extracts DNA/RNA/proteins from the biospecimen. It can be used to streamline the entire pre-analytical workflow, from specimen collection (immediately arrests gene expression and stabilizes the DNA/RNA/proteins) to DNA/RNA/protein extraction (does not use any other DNA, RNA, and protein extraction kits). Therefore, it reduces biospecimen pre-analytical variability, prevents sample cross-contamination, increases detection specificity, and improves data reproducibility and reliability.
Safeguard biospecimens during their transportation and storage
AquaPreserve-stabilized specimens may be stored at 4 and 22 °C for weeks without causing RNA degradation. AquaPreserve serves a safeguard against unexpected cold-chain disasters during shipping and storage, and makes it feasible to repeatedly thaw the frozen the specimens for QC/QA and for distribution to researchers.
Maximize the scientific value of the biospecimens
Presently you probably only store cellular DNA extracted from fresh blood and you don't have a reliable method to extract RNA from frozen whole blood collected in common anticoagulants (not in specialized PAXgene or Tempus RNA tubes). With AquaPreserve, you not only can extract blood DNA but also blood RNA, not only cellular DNA/RNA/proteins but also cell-free circulating DNA/RNA/proteins, from not only fresh but also archived frozen whole blood. It allows you to do more researches and generate more data previously impossible from your precious specimens.
Reduce the overall cost of biobanking
AquaPreserve enables the freezing of leftover blood samples obtained from routine testing for later DNA/RNA/protein extraction, therefore, opens up an unlimited and inexpensive biospecimen resource. AquaPreserve-stabilized the blood biospecimens may be shipped and stored at 4 or 22 °C for years without causing DNA degradation, therefore, reducing the energy consumption. Additionally, AquaPreserve extracts DNA/RNA/proteins from the specimens without needing other DNA, RNA, and protein extraction kits.
Comparison of Techniques for Biospecimen Preservation
Frozen human ACD blood was thawed in 1 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.
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 2 weeks, 22 °C for 7 days.). Centrifuge at 14,000xg for 10 min to pellet the proteins (save the pellet for protein recovery with ProMelt (#1115), if desired).
3. Pellet the DNA/RNA
Pour the supernatant (~0.7 ml) to a 1.5-ml microfuge tube preloaded with 0.8 vol (~0.56 ml) of isopropanol and vortex to mix. Centrifuge at 14,000 xg for 10 min to pellet the DNA/RNA. Decant to discard the supernatant (or save it for small molecule analysis) 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
* This is the RNA yield from human blood. For mouse blood, RNA yield may be 2500 ng/50 ul blood.
If you need to recover the plasma for other assays or prepare DNA/RNA/proteins from large volume of blood while reducing the consumption of the extraction reagents, you may prepare buffy coat from whole blood or use existing frozen buffy coat samples for DNA/RNA/protein extraction. The protocol below is for processing ~2 ml of whole blood to obtain ~200 ul of buffy coat. If you need to process larger volume of whole blood in the original vacutainer (5-10 ml), you will simply scale up the reagent volumes proportionally.
1. Prepare the buffy coat
Centrifuge 2 ml of anticoagulated whole blood at 300 xg for 10 min at room temperature. Remove some plasma (~0.6-0.7 ml) without disturbing the buffy coat. Set the pipette at 100-ul and carefully suck up the grayish buffy coat while slowly moving the tip across the interface and taking up as little RBC as possible. Transfer the buffy coat to a 1.5-ml microfuge tube. Repeat it by taking 100 ul of plasma just above the interface. The total volume of buffy coat recovered is about 1/10 of the blood volume, that is, ~200 ul.
2. Lyse the blood cells
Add one volume (~200 ul) of AquaPreserve (don’t forget to invert the reagent bottle to mix the reagent well before dispensing) to the buffy coat. Vortex to mix well.
3. Recover the DNA/RNA
Add 0.8 volume (~320 ul) of isopropanol to the cell lysate. Vortex to mix well. Centrifuge at 12,000 xg for 5 min at room temperature to pellet the DNA/RNA. Transfer 0.4 ml protein-containing supernatant to a 2-ml tube for protein recovery. Remove the remaining supernatant from the DNA/RNA pellet as much as possible. Fill up the microfuge tube with 50% isopropanol to rinse the DNA/RNA pellet and quickly decant to discard the isopropanol solution. Repeat the isopropanol rinse once. Tap the tube on a clear paper towel to remove residual isopropanol and leave the tube up side down to air dry the DNA/RNA pellet for 5-10 min. Add 100 ul of deionized water to the pellet, and vortex to suspend the DNA/RNA pellet. Incubate at room temperature for 5-10 min and centrifuge again to pellet any insoluble. Transfer the DNA/RNA solution to a new tube and store at –20 °C.
4. Recover the proteins
Add 4 volume (1.6 ml) of acetone to the isopropanol supernatant obtained after DNA/RNA precipitation. Shake or vortex to mix well. Centrifuge at 12,000 xg for 5 min to pellet the proteins. Decant to discard the supernatant. Immediately add 100 ul of ProMelt (#1150, order separately) to the wet protein pellet. Pipet and vortex to solubilize the proteins. Centrifuge to pellet any insoluble. The protein solution may be loaded directly to SDS-PAGE.
(Note: If the buffy coat contains large amount of RBC, you may need to use 2 volumes of AquaPreserve for the extraction or try various volume of ProSink (#9030) for protein precipitation to reduce hemoglobin contamination of the DNA/RNA pellet.)
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
Kit Contents: The AquaPreserve Kit includes the following items
Ordering: To order, please click the "BUY" button below (order ProSink and ProMelt separately).
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