For the majority of patient-derived xenograft lines in the Mayo Clinic Brain Tumor Patient-Derived Xenograft National Resource, RNA was extracted using the QIAshredder Kit (Qiagen #79656) along with the RNeasy Mini Kit (Qiagen #74106) from triplicate biological replicate flank tumor samples for each PDX line.
RNA is stored at minus 80 degrees Celsius and cDNA is stored at minus 20 degrees Celsius.
cDNA was subsequently synthesized using these reagents:
- Random Primer (Thermo Fisher #48190011)
- PCR Nucleotide Mix (Sigma-Aldrich #11814362001)
- RNasin Ribonuclease Inhibitor (Fisher Scientific #PR-N2111)
- M-MLV Reverse Transcriptase (Fisher Scientific #PR-M1701)
Aliquots of cDNA and RNA also are available. To request aliquots, contact the PDX National Resource.