NGS2-3 - Pool of 45 cDNA clones
Drosophila RNA was isolated from S2 cells, reverse-transcribed and selected cofactors' cDNAs (lacking the STOP codon) were amplified using designed primers containing attB sites for Gateway BP cloning (Invitrogen). The 5' and 3' flanks of resulting entry clones were verified using SANGER sequencing and were then subjected to further validation using deep sequencing. For this, a pool of 5µg containing 100-300 clones was sonicated and subjected to library preparation followed by deep-sequencing.
Research Institute of Molecular Pathology (IMP), V
NGS2-3