Animals from a ry[-] line carrying P{PZ} on a second chromosome CyO balancer chromosome were mated to ry[-] animals carrying P{Δ2-3}99B, a stable source of transposase. Transposition events were identified by appearance of ry[+] animals that were not Curly.

In a screen designed to detect insertions on the free minichromosome Dp(1;f)1187, insertions on the autosomes were also recovered. Parental males carried a P{PZ} insertion on the X chromosome and the transposase-producing P{Δ2-3}99B construct on the third chromosome; they also carried a copy of Dp(1;f)1187. They were crossed to ry[-] females and their progeny (F1) screened for ry[+] males. F1 ry[+] males were mated individually to determine the chromosomal location of the transposed insertion, and stable stocks were established. No more than 2 males from a single parental male were characterized to minimize premeiotic clusters.

An aggregate collection of lines produced in two different laboratories (see below).

Markers in stocks (as originally submitted to the BDSC): cn¹, ry⁵⁰⁶ in the Spradling lethal and sterile lines; ry⁵⁰⁶ in the Rubin ‘r’ lines.

General location of each insertion was determined by in situ hybridization to polytene chromosomes. Insertions that mapped to the same cytological location were tested for complementation. Using an array of characterized autosomal deficiencies, in many cases it was possible to test for complementation with an appropriate deficiency; failure to complement resulted in the line being characterized as "verified." Flanking genomic sequences were isolated by plasmid rescue (primarily) or by inverse PCR. Original collection consisted of 528 validated lines.

459 lines were selected for inclusion in the BDGP Gene Disruption Project 2004 collection.