Gene expression profile detection kit for predicting breast cancer reoccurrence of Chinese population
A technology for detecting kits and gene expression profiles, applied in the field of medical molecular biology, to achieve good clinical application prospects and the effect of preventing recurrence
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[0012] (1) extraction of breast cancer paraffin specimen mRNA: (Ambion company RecoverALL was adopted TM Total Nucleic Acid Isolation Kit)
[0013] 1. Cut 5-6 10μm wax slices with a paraffin microtome and put them in a 1.5mL RNA free centrifuge tube.
[0014] 2. Add 1mL xylene for dewaxing, 50°C water bath, 3-5min, 12000rpm, 2min, discard xylene.
[0015] 3. Add 1mL of 100% ethanol, 12000rpm, 2min, discard the ethanol, repeat the ethanol wash once (if the dewaxing is not complete, add xylene and repeat the above steps).
[0016] 4. After blotted with ethanol, air dry for 20 minutes, add 100 μL of digestion solution and 4 μL of protease.
[0017] 5. After mixing, 50°C, 15min; 80°C, 15min.
[0018] 6. Add 120 μL of Isolation additive and 275 μL of 100% ethanol, mix up and down.
[0019] 7. Put the filter element into the collection tube, add the above mixed solution into the filter element, 10000rpm, 30s.
[0020] 8. After discarding the waste liquid, add 700μL wash1 to the...
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