Figure 1 from FUME-TCRseq Enables Sensitive and Accurate Sequencing of the T-cell Receptor from Limited Input of Degraded RNA

Schematic representation of the FUME-TCRseq methodology, whereby RNA is reverse-transcribed using a UMI-containing primer, followed by multiplex PCR amplification of the CDR3 region using a panel of 38 primers and a second PCR to add sample-specific indexes. SP1 (33bp) and SP2 (24bp) are binding sites for universal Illumina sequencing primers and are used for paired-end sequencing of the amplified TCR and sequencing of the 8bp index 1 (i1) and the 8bp index 2 (i2). P7 (24bp) and P5 (29bp) are flow cell binding sequences for Illumina-sequencing platforms.