Gene discovery using massively parallel pyrosequencing to develop ESTs for the flesh fly Sarcophaga crassipalpis

Table 1 RNA Extraction Strategy.

Pre-adult stages		Adult Stages
Exposed to short days to induce diapause development	Exposed to long days to induce direct development	Non-reproductive Fed only sugar	Reproductive Fed sugar and protein (liver)
First Instar Larvae	First Instar Larvae	Day 1 Males	Day 1 Males
Second Instar Larvae	Second Instar Larvae	Day 1 Females	Day 1 Females
Early Third Instar Larvae	Early Third Instar Larvae	Day 2 Males	Day 2 Males
Late Third Instar Larvae	Late Third Instar Larvae	Day 2 Females	Day 2 Females
Wandering Larvae	Wandering Larvae	Day 3 Males	Day 3 Males
Pupae	Early Diapause Pupae (15 days)	Day 3 Females	Day 3 Females
Early Pharate Adult	Mid-diapause Pupae (30 days)	Day 4 Males	Day 4 Males
Mid-Pharate Adult (red eye stage)	Late Diapause Pupae (60 Days)	Day 4 Females	Day 4 Females
Late Pharate Adult (black bristle stage)		Day 5 Males	Day 5 Males
		Day 5 Females	Day 5 Females
		Day 7 Males	Day 7 Males
		Day 7 Females	Day 7 Females
		Day 9 Males	Day 9 Males
		Day 9 Females	Day 9 Females

Flesh fly life stages that were individually RNA extracted and combined equally prior to cDNA library synthesis and normalization.

ISSN: 1471-2164