title: GSM2256203: ncm_BAC_S2_rep2; Drosophila melanogaster; OTHER
library_construction_protocol: Genomic DNA (source: embryos of the sequenced strain: y; cn bw sp) or BAC DNA was isolated, sheared and size selected (~100-250bp). Following the instructions of NEBNext® Ultra™ II DNA Library Prep Kit for Illumina® (NEB; cat. no. E7645L) ,Illumina NEBnext Multiplexing Adaptors (NEB; cat. no. E7335 or E7500) were ligated and homology arms for In-Fusion® recombination were added by PCR, followed by recombination into the STAP-seq vectors. To construct the STAP-seq vector, the sequence between BglII and FseI from the pGL3-Promoter backbone (Promega; cat. no. E1751) was replaceda ccdB suicide gene flanked by homology arms (used for cloning the promoter candidates during library generation), an intron (mhc16), an ORF (truncated sgGFP, Qbiogene, Inc), followed by the pGL3's SV40 late polyA-signal. The enhancers were cloned between the KpnI and BglII sites. The control screens were performed with a vector not harboring an enhancer. The In-Fusion® reactions were transformed (MegaX DH10B; Invitrogen), grown in liquid culture and plasmids were isolated.