Single-cell genomic & transcriptomic analyses of longitudinal samples of patients with RichterSyndrome reveal the presence and dynamics of clones driving transformation from chronic lymphocytic leukemia years before clinical manifestation. FerranNadeu
with minor modifications. Single cells were sorted into 96-well plates containing the lysis buffer . Reverse transcription was performed using SuperScript II in the presence of 1 μM oligo-dT30VN , 1 μM template-switching oligonucleotides and 1 M betaine . cDNA was amplified using the KAPA Hifi Hotstart ReadyMix and IS PCR primer , with 25 cycles of amplification.
-negative live individual cells were sorted with a BD FACSAria Fusion Flow cytometer in 1× PBS supplemented with 0.05% BSA. After FACS, cells were partitioned into Gel Bead In Emulsions by using the Chromium Controller system aiming at a Target Cell Recovery of 5,000 total cells. Sequencing libraries were prepared using the v.3.1 single-cell 3′ mRNA kit . After GEM-RT cleanup, cDNAs were amplified during 14 cycles.
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