title: H.melpomene malleti wing transcriptome
design: Developing wings were dissected from multiple individuals from a laboratory stock of Heliconius melpomene malleti at developmental stages from late larval through to mid-pupal stage.  mRNA was pooled by race according to the following proportions: 40% mid 5th instar larvae, 20% late 5th instar, 20% early pupae and 20% pre-ommochrome pupae. First strand cDNA was synthesised using the Creator SMART cDNA Library Construction Kit (Clontech). Double stranded cDNA (ds-cDNA) was synthesised and normalised following the Trimmer-Direct Kit for cDNA normalization (Evrogen) guidelines.  The optimal condition for ds-cDNA synthesis for each single stranded cDNA template was empirically determined: reaction mixes were subjected to a recommended range of thermocycle numbers and their products checked by electrophoresis (1 % agarose gel). The optimal cycle number was defined as maximum cycle number without signs of overcycling. Ds-cDNA was then purified and concentrated using the DNA Clean and Concentrator kit (Zymogen).