GSM4276514: PfArp4_R_rep2_RNAseq; Plasmodium falciparum; RNA-Seq
The experiment began to collect the highly synchronized Plasmodium, centrifuge and remove the supernatant, then the cells was lysed by adding 1:5 triazole. Next to depolymerize the proteins and nucleic acids in the cells to release them, and extract RNA according to reagent specification of Direct-zol RNA MiniPrep (R2052). The resullts of RNA extraction were verified on a 2% agarose gel. The mRNA was enriched with magnetic beads with Oligo (dT). It has been optimized to isolate mature mRNA from total RNA by capturing twice using the KAPA mRNA Capture Beads. This experiment was performed using the KAPA Stranded mRNA-seq Kit. Here, the required fragment size for the experiment is 300-400 bp. It carried out the fragmentation and priming program as follows: 85°C 6min. In particular, some changes to the library amplification procedure have been made as follows: initial 98°C 45sec 1cycle; denaturation 98°C 15sec 14cycles; annealing 65°C 30sec 14cycles; extension 65°C 30sec 14cycles; final extension 65°C 5min 1cycle; hold 10°C 1cycle. Resuspend the dried beads in 22µl of RNase-free water to obtain the final library building results. The library were sequenced on an Illumina HiSeq Xten systerm with PE150 strategy.
GEO Accession
GSM4276514