Ezway taq pcr mastermix

Validation of gene fusion transcripts were performed by reverse transcription polymerase chain reaction (RT-PCR) assays. Total RNA was extracted from the tissues by AllPrep DNA/RNA Mini kit according to the manufacturer’s instructions (Qiagen). Total RNA (1 μg) was reverse transcribed to synthesize template cDNA by a random primer using the SuperScriptIII First-Strand System(Life Technologies), and 20 μl of synthesized cDNA was diluted 5 fold with DW. For RT-PCR, EzWay Taq PCR MasterMix (Komabiotech, KOREA) and 5 μl of synthesized cDNA template was used. Thermal cycling was carried out under the following conditions: 1 min at 95°C followed by 30 cycles of 30 sec at 95°C, 30 sec at 56°C, 30 sec at 72°C. The primer pairs used in this experiment were designed to make the amplification product including the breakpoints of the fusion genes. PCR products were analyzed by agarose gel electrophoresis. The primers were summarized in Supplementary Table 6.

For validation of fusion transcripts and RT-PCR assays were performed. Total RNA was extracted from the tissues by AllPrep DNA/RNA Mini kit according to the manufacturer’s instructions (Qiagen). The total RNA (0.5 μg) was reverse transcribed to synthesize template cDNA by a random primer using the SuperScriptIII First-Strand System(Life Technologies), and 20 μl synthesized cDNA was diluted 10 times with DW. For RT-PCR, EzWay Taq PCR MasterMix (Komabiotech, KOREA) and 5 μl synthesized cDNA as template were used. Thermal cycling was carried out under the following conditions: 1 min at 95°C followed by 30 cycles of 30 sec at 95°C, 30 sec at 55°C, 30 sec at 72°C. The primer pairs used in this experiment were designed to make the amplification product including the breakpoints of the fusion genes. PCR products were analyzed by agarose gel electrophoresis. The primers were summarized in Supplementary Table 11.