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Interestingly, wAnaINF from D. ananassae strain HNL0501 had the lowest wAna originating reads and normalized average read depth (table 1).
In addition, a recent RNA sequencing study detected only a few WLGT originating reads out of the total Drosophila ananassae RNA pool (Kumar et al. 2012), suggesting that the observed gene expression could be background transcriptional noise.
Whole adult flies representing each of the 15 strains of D. ananassae that were previously screened for the presence of wAnaINF and wAnaITG genomes (Choi and Aquadro 2014) were genome sequenced, and all wAna originating reads were computationally extracted using the reference wRi genome (alignment statistics are shown in table 1).
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The paired-end protocol was used and originated reads of approximately 100 bp for the sequencing of cDNA libraries.
As the DNemulator package generates read names that include the position from which the read originates, reads in a pair do not have identical names, which RetroSeq and Jitterbug both rely on.
The small 9613 bp contig from the PacBio assembly was found to match the 7.9 Mb contig from position 5938858 to 5929122 (reverse complement) but with only 92 % identity, possibly indicating that it originated from reads with accumulated errors.
Mapping locations indicate transcripts of origin, and the number of reads originating from a given transcript inform on how much of the transcript was present in the sample.
For this class, the PCR products and sequence reads originating from the positive versus negative strand of the original recombinant molecule were identical.
After assembly, transcripts with length greater than 90 base pairs (bp) were subjected to expression pattern examination as follows: the numbers of reads originating from the different cDNA libraries for each transcript was recorded, and transcripts with reads originating from only one stage (or a set of stages) were designated to be specific to that stage (or that set of stages).
Furthermore, we carefully inspected reads originating from tRNAs and snoRNAs, since recent reports have highlighted a novel class of small RNAs originating from these structural RNAs [ 8- 12].
Second, reads originating from the 3' end of exons would not be aligned to the genome, since the reads originate from ligations of two exons and not from consecutive genomic regions.
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