Fig. 5

In vitro validation techniques. a Example of PCR validation on deletion (Del_218). Lane 1 is the DNA marker; Lane 2 is the test sample; Lane 3 is the reference control; Lane 4 is the no template control (NTC). The test sample has a deletion in the target position which makes its amplification PCR size smaller than reference control. b Example of ddPCR validation on duplication (Dup_1158). NA19239 is the test sample. NA10851 and NA12878 are reference controls. NTC is the no template control. Duplicates were run to avoid random experimental error in all ddPCR experiments. NA19239 has an amplification compared to the control. This candidate has been validated. c Example of Sanger sequencing validation on Inversion (Inv_190). d Sanger sequencing chromatogram to identify inversion. The arrows indicate the breakpoints from where the sequences between test sample and control become different with each other. The yellow arrows indicated the predicted left and right breakpoints using FusorSV algorithm and the blue arrows indicated the sequenced breakpoints by Sanger sequencing. Reference: reference genomic sequences (GRCh37/hg19 Assembly) extracted from UCHC Genome Browser; Inversion_Ref: predicted inversion sequences by FusorSV; Inversion_inverted: inverted inversion sequences; Test_NA12878: nucleotide sequences from Sanger sequencing on test sample NA12878. Control_NA10851: nucleotide sequences from Sanger sequencing on control sample NA10851