Sanger sequence results from genomic DNA, with primer FGFR2-G3BP2_Fwd
FGFR2-G3BP2 and SAMD13-FGFR2 genomic breakpoints were validated by PCR and Sanger sequencing. PDC-DUC18828 (1x106 cells) were dissociated and resuspended in cell media, centrifuged and washed in PBS, and then pelleted by centrifugation. Genomic DNA was then extracted using Qiagen Blood and Cell Culture DNA Kits (Cat #13323) according to the manufacturers protocol.  PCR was performed on purified DNA using GoTaq (Promega) PCR master mix using primer pairs FGFR2_SAMD13_F/ FGFR2_SAMD13_R and  FGFR2_G3BP2_F/ FGFR2_G3BP2_R at an annealing temperature of 55C.  Crude PCR fragments were purified using Ampure XP beads (Beckman Coulter) according to the manufacturers protocol.  Purified PCR fragment was subjected to Sanger DNA sequencing with the same forward and reverse primers used for PCR. DNA sequencing was performed using BigDye v1.1 and electrophoresed on an ABI 3730xl instrument using POP7 according to the manufacturers protocol.