single cell splicing sequencing of Human immunodeficiency virus 1:LR3_AS_SAHA_R2_0.01ng
I added a UMI to each viral mRNA molecule immediately downstream the barcode sequence by reverse transcription. Then I amplified all types of the whole HIV-1 transcripts. The PCR product was enzymatically digested and circularized to bring the 5 UTR and the barcode region close. Lastly, I used 3 different sets of primers to amplify the barcode and 5 UTR region, they targeted different splicing families.