GSM7767315: Barcoding_library_of_HSCs; Mus musculus; OTHER
For targeted CARLIN amplification based on scRNA-seq cDNA, 5 ng amplified cDNA was used for the first PCR reaction (10X-CARLIN_1- bio, P5-PR1; 9 cycles). After cleanup with 0.8X SPRI beads (Beckman Coulter), the PCR products were purified using Dynabeads kilobase BINDER Kit (Invitrogen). The cleanup products were then used for second PCR reaction (NGS1_Bulk_R, P5-PR1; 9 cycles). After Dynabead removal, the PCR products were purified with 0.8X SPRI beads for the indexing PCR (SI primer, Chromium i7 sample primer; 8 cycles). The libraries were sequenced on Illumina MiSeq using paired-end 500 cycles v2 kits (Read 1: 28 cycles; Index Read (i7): 8 cycles; Read 2: 350 cycles) with 10% PhiX sequencing control v3 (Illumina)