Skip to main content
. 2022 Aug 18;4(1):vdac131. doi: 10.1093/noajnl/vdac131

Figure 3.

Figure 3

Detection of the hybrid chromosome containing the 1p/19q arms. (A) Schematic of the PCR assay that specifically detects the hybrid chromosome containing the 1p/19q arms (the 1p/19q product). To ensure reaction specificity, primer pairs recognizing 1p and 19q (arrows) and an internal Taqman probe recognizing 19q sequences have been used. INDELs, insertions and deletions of several nucleotides at the DSB repair site. Note that the internal probe is designed to recognize sequences outside the anticipated Cas9-cutting site and INDEL region. (B) Droplet digital PCR (ddPCR) analysis of the 1p/19q product in HEK 293T cells collected on different days after CRISPR/Cas9 introduction. CCR5, internal control to normalize the copy number of input genomic DNA. The percentage shows the fraction of 1p/19q positive droplets of each sample. (C) Electrophoresis of the 1p/19q product. Arrow, the expected PCR product. DNA ladder, 1KB Plus (NEB #N3200S). (D-E) Automatic sequencing analysis of the 1p/19q product using (D) the 1p primer and (E) the 19q primer. Red arrowheads, Cas9 cutting sites.