Xp dna beads

Single-cell RNA-seq library construction was performed as previously described (Cui et al., 2019 (link); Lu et al., 2019 (link)). This modified method was based on the STRT-seq and Smart-seq2 methods. Briefly, the amplified cDNA bearing different cell barcode primers were pooled and purified with 0.8× XP DNA beads (Beckman) twice and then amplified for four cycles with primers with the Illumina index sequence and a biotin modification. After purification with 0.8× XP DNA beads once (Beckman), the DNA was sheared to approximately 300 bp by ultrasonicator (Covaris S2), followed by Dynabeads MyOne Streptavidin C1 beads (Invitrogen) to capture the 3ʹ cDNAs. Thereafter, the library was constructed using a Kapa Hyper Prep Kit (Kapa Biosystems) and the cleaned library was sequenced on an Illumina HiSeq 4000 platform with 150-bp paired-end read length.

Single-cell RNA-seq library construction was performed as previously described (Cui et al., 2019 (link); Lu et al., 2019 (link)). This modified method was based on the STRT-seq and Smart-seq2 methods. Briefly, the amplified cDNA bearing different cell barcode primers were pooled and purified with 0.8× XP DNA beads (Beckman) twice and then amplified for four cycles with primers with the Illumina index sequence and a biotin modification. After purification with 0.8× XP DNA beads once (Beckman), the DNA was sheared to approximately 300 bp by ultrasonicator (Covaris S2), followed by Dynabeads MyOne Streptavidin C1 beads (Invitrogen) to capture the 3ʹ cDNAs. Thereafter, the library was constructed using a Kapa Hyper Prep Kit (Kapa Biosystems) and the cleaned library was sequenced on an Illumina HiSeq 4000 platform with 150-bp paired-end read length.