RNA-Seq of Drosophila melanogaster: Dscam1-L RT-PCR; 16-20 hour embryos
Amplification from cDNA of a region encompassing Dscam1 exon 16 until within the extended 3  UTR was performed using the following primers (forward: 5  - TTTCTGTTGGTGCTGATATTGCCGAATACGACTTTGCCACCT - 3 ; reverse: 5  - ACTTGCCTGTCGCTCTATCTTCTCTGTAGCTCCATTGCATCG - 3 ). The PCR product was purified using AMPure XP beads (A63881, Beckman Coulter Life Science), and then used as a template for PCR amplification using nanopore barcodes primers (SQK-LSK108 Ligation Sequencing Kit 1D, and BC001, BC002, BC010 in EXP-PBC001 PCR Barcoding Kit I, from Oxford Nanopore Technologies) to obtain barcoded amplicons. Gel extracted PCR products (1 g in total) was used for Nanopore sequencing with FLO-MIN 107 flowcell.