GSM4627568: tgrB1_r1_hr20; Dictyostelium discoideum; RNA-Seq
At each time point, we harvested the cells directly into 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 of all samples. 434 individual libraries were separately prepared and divided to 22 sets. We added a unique barcode to each library in each set at the final step of PCR amplification. We pooled equal amounts of DNA from each library and sequenced those pools by Illumina HiSeq2500 (single-end sequencing, read length = 50 or 100 bases) or (paired-end sequencing, read length = 93 or 100 bases) or by NovaSeq6000 (paired-end sequencing, read length = 150 bases).
GEO Accession
GSM4627568