Description
General description
For simultaneous cell lysis and stabilization of nucleic acids in blood or bone marrow samples.
Application
The RNA/DNA Stabilization Reagent for Blood/Bone Marrow is intended for general laboratory use and aids in the collection of samples for nucleic-acid isolation. It is able to lyse cells and simultaneously inactivate nucleases when added to blood or bone-marrow samples. In addition to providing safe transport from the collection site to the laboratory, the reagent also serves as an effective long-term preserving agent, protecting nucleic acids at –15 to –25°C until future analysis.
The easy-to-use reagent increases the accuracy of nucleic-acid detection by ensuring complete and immediate inactivation of nucleases. At the time of collection, one volume of blood sample or bone-marrow aspirate is added to 10 volumes of reagent, thus minimal reagent calculations are needed.
Preparation Note
The RNA/DNA Stabilization Reagent for Blood/Bone Marrow contains a strong protein denaturant (e.g., guanidinium thiocyanate), detergents, and reducing agents to accomplish instantaneous cell lysis and nuclease inactivation. The nucleic acids are stabilized, and ready for transport back to the laboratory for isolation with the mRNA Isolation Kit for Blood/Bone Marrow.
Lysates can be immediately used, stored for one day at +2 to +8°C, or stored for up to 1 year at –15 to –25°C. Five hundred milliliters is sufficient to stabilize a total of 50 mL of whole blood or bone-marrow aspirate.
Analysis Note
The reagent is function-tested in the following model system: the detection of the equivalent of five melanoma cells in a volume of 5 mL of normal human blood.
The recovery of tyrosinase mRNA is detected by RT-PCR after purification of 5 pg of MelJu cell mRNA, added to a lysate prepared of 5 mL of fresh normal human blood, and stabilized with the RNA/DNA Stabilization Reagent for Blood/Bone Marrow. The mRNA is purified from the stabilized lysate with the mRNA Isolation Kit for Blood/ Bone Marrow.
Note: 5 pg of MelJu cell mRNA is equivalent to about five melanoma cells.
Other Notes
For life science research only. Not for use in diagnostic procedures.
