RNA was then extracted from the sorted cells using the Qiagen RNeasy kit, treated with DNase I, and cDNA was synthesized using the Bio-Rad iScript cDNA synthesis kit.
Total RNA was purified with a Qiagen Extraction Kit, treated with DNase I, and 1 µg of total RNA was reverse-transcribed using Superscript II (Invitrogen).
Total RNA was extracted using RNeasy® RNA purification kit, treated with DNase-I and RNA content was determined by measuring the absorbance at 260 nm.
Where indicated, total cellular RNA was extracted from tumors using RNeasy Mini Kit, treated with DNase I, and converted to cDNA with the RT First Strand Kit according to the manufacturer's instructions.
RNA were extracted [ 56] with the RNeasy kit from Qiagen (DNA microarrays kit) and treated with RNase-free DNase I (Roche).
The total RNA from the chondrocytes (negative control) was extracted using an RNeasy Mini Kit (Qiagen), treated with DNAse I (Invitrogen), and reverse transcribed with a High Capacity cDNA Archive Kit (Applied Biosystems).
Total RNA was extracted with the RNeasy Mini Kit (Qiagen), treated with DNAse (recombinant DNase, Roche) and subsequently transcribed into cDNA (Transcriptor First Strand cDNA Synthesis Kit, Roche Diagnostics).
RNA was isolated from the ground cartilage using the RNeasy kit (Qiagen), treated with Turbo DNase (Ambion) and then reverse transcribed using the SuperScript First-Strand cDNA synthesis kit (Invitrogen).
The difference in median OS times between MCTs with c-KIT mutations treated with local therapy alone and those treated with vinblastine and prednisone suggests that MCTs with c-KIT mutations also might benefit from postoperative chemotherapy with vinblastine and prednisone.
Total RNA from ZR-75-1 cells was purified using the GenEluteTM Mammalian total RNA miniprep kit (Sigma), treated with amplification grade DNase I (Sigma), and measured using a NanoDropTM spectrophotometer.
Total RNA was extracted using the RNeasy mini kit and treated with DNaseI (Qiagen).
