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Analysis Note
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.
Application
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.
General description
Other Notes
Preparation Note
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.
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Danger
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Hazard Classifications
Acute Tox. 4 Oral - Aquatic Chronic 3 - Eye Dam. 1 - Skin Corr. 1C
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Storage Class
12 - Non Combustible Liquids
wgk
WGK 2
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No data available
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No data available
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A poly(A)-mRNA purification procedure applying Boehringer Mannheim's new RNA/ DNA Stabilization Reagent for Blood/Bone Marrow and the new mRNA Isolation Kit for Blood/Bone Marrow, was tested to prove its applicability for the detection of disseminated tumor cells from peripheral blood, by use of the poly(A)-mRNA fraction in combination with a highly sensitive RT-PCR procedure. In order to establish the sensitivity of the technique, peripheral blood samples from healthy donors were spiked with defined numbers of melanoma cells from an established tumor cell line.
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