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Fig. 1 | Genome Biology

Fig. 1

From: Vex-seq: high-throughput identification of the impact of genetic variation on pre-mRNA splicing efficiency

Fig. 1

Assembly of test exon and experimental design. a The test exon and flanking introns are subcloned into a reporter plasmid in a two-step process, such that the barcode designating the sequence is near the end of the transcript. Once these plasmids are transfected into cultured cells, a transcript will be produced that may not contain the variant itself, but does contain the barcode (b) uniquely associated with the variant tested. A ten-nucleotide UMI (N10) is attached during the reverse transcription step to collapse PCR duplicates downstream. Illumina flow cell binding sequences (FC) and indexes (I1 and I2) are attached via primers during PCR and the resulting DNA is sequenced on a MiSeq platform. b Data analysis pipeline for splicing results

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