Rupi_Control_Condition_Rep_A
ProtoScript II First Strand cDNA Synthesis Kit (New England Biolabs, E6560L) was used to generate first strand cDNA using a reverse primer in the 3UTR, TGGTTATAACAAATTTACCCCCACCA. Then, the cDNA was used as a template in four independent Q5 PCR reactions with 25 cycles. Primers for amplifying the replication protein region were TCAAAGCCAACCCAAATTATGCT (forward) and TGGTTATAACAAATTTACCCCCACCA (reverse). Sequencing libraries for the input plasmid library were prepared as above, beginning with the PCR step. Gel purified amplicons were used as input for Illumina sequencing library preparation using the Twist Biosciences Enzymatic Fragmentation 2.0 kit with Universal Adapters (Twist Biosciences, 104207) with 180-220 bp target fragment sizes. Illumina libraries were pooled (6 samples per run) and sequenced with a NextSeq 2000 XLEAP P2 flow cell, 300 cycle paired-end kit (Illumina, 20100985).
SUB15938590
Rupi_Control_Condition_Rep_A