title: GSM4254229: Cmi RNAi depletion BG larvae; Drosophila melanogaster; RNA-Seq
library_construction_protocol: Animals (>20 of each genotype) were collected, and RNA extracted using RNAqueous (Invitrogen). Samples were DNAseI treated, RNAs selected by oligo dT and fragmented. Then cDNA is synthesized  using the mRNA fragments as templates. Short fragments are purified and resolved with EB buffer for end reparation and single nucleotide A (adenine) addition. After that, the short  fragments are connected with adapters. After agarose gel electrophoresis, the suitable fragments are selected for the PCR amplification as templates. During the QC steps, Agilent  2100 Bioanaylzer and ABI StepOnePlus Real-Time PCR System are used in quantification and qualification of the sample library. At last, the library could be sequenced using  Illumina HiSeqTM 2000. Primary sequencing data that produced by Illumina HiSeqTM 2000, called as raw reads, is subjected to quality control (QC) that determine if a resequencing step is needed. After QC,  raw reads are filtered into clean reads which will be aligned to the reference sequences with SOAPaligner/SOAP2[ 1]. QC of alignment is performed to determine if resequencing is  needed. The alignment data is utilized to calculate distribution of reads on reference genes and perform coverage analysis. Libraries were prepared according to Illumina's instructions and performed by Beijing Genomics
attributes.tag: GEO Accession
attributes.value: GSM4254229