GSM9112730: Psc-Su(z)2-KO-N1 , TRIP-normalization library, rep1; Drosophila melanogaster; OTHER
TRIP-mapping libraries: Genomic DNA was MboI-digested, circularized, and used for three rounds of inverse PCR to amplify TRIP barcodes and insertion sites. Libraries were cleaned, quantified, and pooled at 10?nM. TRIP-normalization and TRIP-cDNA libraries: Genomic DNA or 10% of cDNA was PCR-amplified in two rounds to introduce sequencing adaptors. Libraries were cleaned, quantified, and pooled. Fractions of TRIP-mapping, TRIP-normalization and TRIP-cDNA libraries were combined into a single 10nM pool at ratios 1:2:2. The pool was sequenced on 1 lane of 10B flow cell using the NovaSeq X Plus system and XLEAP-SBS sequencing chemistry.