title: GSM8583767: AX4_20hr : replicate 2; Dictyostelium discoideum; RNA-Seq
library_construction_protocol: At each time point, we harvested the cells, resuspended with 1 mL of Trizol® (life technologies, CA, USA) and extracted total RNA according to the manufacturer's recommended protocol. We performed two rounds of poly-A selection and fragmented 100 ng of the resulting mRNA into approximately 200 bases fragments. We prepared cDNA and the second strand. We prepared Illumina multiplexed libraries. We prepared 108 individual libraries separately (one from each sample), three categories and added a unique barcode to each library in each category at the final step of PCR amplification. We pooled equal amounts of DNA from each library in each category and sequenced one pool per lane of a flow cell on Illumina NovaSeq6000 using the manufacturer's recommended pipeline (PE, read length = 151 bases). Since the illumina sequence reads for rapgapB- and rapgapB- tgrB1- strains were low, they were sequenced twice. Therefore, we obtained 4 fastq file for each of these samples. We combined those fastq file to analyze them.
identifiers.namespace: GEO
identifiers.id: GSM8583767_r1