Transcriptional profiling of Drosophila intestine in aging and identification of genes

The fruit flies used in this study were maintained at a constant temperature of 25 degrees Celsius on a standard cornmeal medium. The specific fly strain utilized was w1118, which were black-bellied female fruit flies. Following hatching, the flies were collected and aged at the same temperature of 25 degrees Celsius. Intestinal dissections were performed on aged female flies at various time points, specifically on days 3, 15, 30, 40, and 50, in chilled phosphate-buffered saline (PBS).Total RNA was then isolated from these samples using the Trizol method, incorporating a DNase treatment to remove any potential DNA contamination. This process was carried out at the Beijing Genomics Institute (BGI Co., Ltd., China). To verify the quality and integrity of the extracted RNA, an Agilent 2100 Bioanalyzer (Agilent Technologies, Palo Alto, CA) was employed.For the purpose of mRNA sequencing, libraries were constructed using the Illumina TruSeq Stranded mRNA Sample Prep Kit (Illumina, Cat. No. RS-122-2101). Subsequently, paired-end sequencing with a read length of 50 base pairs was performed on an Illumina HiSeq 2000 system. Each RNA-Seq experiment included three biological replicates, with each replicate containing approximately 200 fly guts.