Crosses and age group regarding Recombinant Complex Intercross Lines (RAILs)
A number of affairs can be determine recombination inside the D. melanogaster including ladies age, temperature, quantity of matings otherwise dinner –. To reduce the consequences ones facts, most of the crosses and you may stresses have been maintained in one temperatures (23C) around a good step one2 hours. light/dark period, which have lingering fly density in half pint bottle into the practical corn-buffet media. Recombination was large getting younger (1–two days) and you will old (>several days) people . For everyone crosses, adult challenges were entered in both instructions in order to average away you can maternal effects into the recombination prices.
Thus all crosses was through with flies out-of similar years, having 6 time dated virgin flies permitted to lover and you will set eggs to possess a time period of thirty-six–2 days, right after which the mother and father have been thrown away and kiddies was in fact desired so you can emerge
To maximize the number of recombination events per fly genotyped, we generated recombinant advanced intercross lines (RAIL). To generate RAILs by sibling mating and minimize homozygosity, we crossed five groups of forty F1 flies, twenty males and twenty females, randomly collected from the previous generation and put together in new bottles. The F1 flies were also allowed to mate for 1.5–2 days after which they were discarded and F2 offspring were allowed to emerge. To generate the next generation, flies from all bottles were randomly mixed to create the next groups of twenty males and twenty females. (mehr …)