The TruSight Tumor 15 Kit has been optimized for a specific DNA input amount. Quantify the input DNA before beginning the protocol.
- 20 ng total (10 ng for Mix A and Mix B per sample) human genomic DNA (gDNA) input at a minimum concentration of 2 ng/μl.
- Use a fluorometric quantification method that uses dsDNA binding dyes such as AccuClear (recommended), Qubit, or PicoGreen.
- Dilute starting material in RNase/DNase-free water.
Reference Samples [Optional]
- [Optional] Use a characterized reference material when running the library preparation, such as HorizonDx HD701 or HD200 (requires DNA extraction), or AcroMetrix Oncology Hotspot Control.
- [Optional] Use 5 μl RNase/DNase-free water as a no template control for PCR amplification. Do not sequence the no template control.
Note: Running a reference sample or no template control reduces the total number of unknown samples that can be processed with each library prep kit.
FFPE DNA Extraction
Use the following recommendations to extract DNA from FFPE tissue:
- Use at least 140 mm2 nonmelanoma tissue with at least 30% tumor.
- Use 1 of the following kits to extract DNA from FFPE tissue:
- AllPrep DNA/RNA FFPE Kit (QIAGEN) (Recommended)
- QIAamp DSP DNA FFPE Tissue Kit (QIAGEN)
- ReliaPrep FFPE gDNA Miniprep System (Promega)