Single cell amplicon-seq of CRISPRed NAMPT gene locus at day 14 (rep2)
The DNA samples prepared with the Fluidigm C1 platform were collected and the NAMPT locus was then amplified for each sample using the QIAGEN Multiplex PCR kit (cat. 206145) together with the following 8 base pair barcoded primers:     NAMPT_FWD2_bc_A_GATCTCTC       GATCTCTCcccgctcctcctcatctg  NAMPT_FWD2_bc_B_ATATCGCT       ATATCGCTcccgctcctcctcatctg  NAMPT_FWD2_bc_C_TAGAGATA       TAGAGATAcccgctcctcctcatctg  NAMPT_FWD2_bc_D_AGCGATAG       AGCGATAGcccgctcctcctcatctg  NAMPT_FWD2_bc_E_TCGAGCGA       TCGAGCGAcccgctcctcctcatctg  NAMPT_FWD2_bc_F_CTAGATAT       CTAGATATcccgctcctcctcatctg  NAMPT_FWD2_bc_G_GATCGCGA       GATCGCGAcccgctcctcctcatctg  NAMPT_FWD2_bc_H_CGAGCGAG       CGAGCGAGcccgctcctcctcatctg     NAMPT_REV2_bc_1_GCGCTCGA       GCGCTCGAccgaccgagcagtgactta  NAMPT_REV2_bc_2_AGATAGAG       AGATAGAGccgaccgagcagtgactta  NAMPT_REV2_bc_3_TATCTCTC       TATCTCTCccgaccgagcagtgactta  NAMPT_REV2_bc_4_CTATCTAG       CTATCTAGccgaccgagcagtgactta  NAMPT_REV2_bc_5_GAGCGATC       GAGCGATCccgaccgagcagtgactta  NAMPT_REV2_bc_6_ATAGAGCG       ATAGAGCGccgaccgagcagtgactta  NAMPT_REV2_bc_7_GCTATCGC       GCTATCGCccgaccgagcagtgactta  NAMPT_REV2_bc_8_CGCTCTCG       CGCTCTCGccgaccgagcagtgactta  NAMPT_REV2_bc_9_TCGCTCTA       TCGCTCTAccgaccgagcagtgactta  NAMPT_REV2_bc_10_CGCGATCT      CGCGATCTccgaccgagcagtgactta  NAMPT_REV2_bc_11_TCTCGATC      TCTCGATCccgaccgagcagtgactta  NAMPT_REV2_bc_12_AGAGATCG      AGAGATCGccgaccgagcagtgactta     The combination of these primers provides 96 uniquely barcoded PCR products, one for each Fluidigm 96 IFC single cell DNA sample. The quality of the 96 PCR products was assessed on the TapeStation 4200 platform (Agilent) using a D1000 kit (Agilent, cat.5067-5582). The 96 amplicons were then pooled together at equal volumes and sequencing libraries were prepared with the Ovation Low complexity Sequencing System starting from each individual pool (NuGEN, cat.9092-256). Libraries were sequenced in paired-end mode, at a read length of 2 × 200 bp, using the MiSeq platform (Illumina).
SUB2124680
LIB06050973