Transcription profile of Dirofilaria immitis
Transcription profile of Dirofilaria immitis
RNA was extracted from the worms using the protocol described in Qiagen miRNeasy Mini Handbook October 2007. Several steps were carried out to convert total RNA into a library of template molecules suitable for high throughput DNA sequencing.  The first step involves purifying the poly-A containing mRNA molecules using poly-T oligo-attached magnetic beads.  Following purification, the mRNA is fragmented into small pieces using divalent cations under elevated temperature. Then, the cleaved RNA fragments are copied into first strand cDNA using reverse transcriptase and a high concentration of random hexamer primers.  This is followed by second strand cDNA synthesis using DNA Polymerase I and RNaseH.  Finally, the short cDNA fragments are prepared for sequencing on the Illumina Genome Analyzer IIx.
Experimental Factor: SEX
male