GSM3096992: Ad_WGBS; Trichinella pseudospiralis; Bisulfite-Seq
Prior to library construction, 5 µg of genomic DNA spiked with 25 ng unmethylated  cl857  Sam7 Lambda DNA (Promega, Madison, WI) was fragmented using a Covarias sonication system to a mean size of approximately 200 bp. After fragmentation, libraries were constructed according to the Illumina Pair-End protocol with some modifications. cDNA libraries were prepared  according  to  the  manufacturer's  instructions  (Illumina).  The poly(A)-containing mRNA molecules were purified using Oligo(dT) Beads (Illumina)from 20  g of total RNA from each sample. Tris-HCl (10 mM) was used to elute the mRNA from the magnetic beads. To avoid priming bias when synthesizing cDNA, mRNA was fragmented before the cDNA synthesis. Fragmentation was performedusing divalent cations at an elevated temperature. The cleaved mRNA fragments were converted into double-stranded cDNA using SuperScript II, RNaseH and DNA Pol I, primed by random primers. The resulting cDNA was purified using the QIAquick PCR  Purification  Kit  (Qiagen).
GEO Accession
GSM3096992