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Clones which were not caught by any of these filters were mapped on the mouse genome (release UCSC mm5 [2]) using the sim4 [37] program.
Mouse and human DNA sequences were obtained from the University California Santa Cruz Genome Browser (mouse genome, release Feb 2007; human genome, release June 2007).
Novoalign (Novocraft) was used to align against NCBI mouse genome release 36 (UCSC Feb. 2006 release [mm8]) and a collection of splice junctions generated from Refseq genes [ 28], ENSEMBL genes [ 29], and UCSC Known Genes [ 30].
We used human genome release 36.3, zebrafish genome release 10/06/2008 (International Human Genome Sequencing Consortium), fruit fly genome release 5 (The FlyBase Consortium/Berkeley Drosophila Genome Project) and mouse genome release 37 (Mouse Genome Sequencing Consortium).
The sequence reads were aligned to the mouse genome (release version mm9) using the bowtie aligner (version 0.12.7) (bowtie-bio.sourceforge.net/index.shtml) [19], which was set up to report only uniquely aligning reads.
Likewise, we updated the mouse scores using: (i) 3′ UTRs derived from 19,699 mouse protein-coding genes annotated in Ensembl 77 (Flicek et al., 2014), and (ii) 3′-UTR MSAs across 52 vertebrate species derived from the 60-way multiz alignments in the UCSC genome browser, which used the mouse genome release mm10 as a reference species (Kent et al., 2002; Karolchik et al., 2014).
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We aligned the reads against the reference mouse genome (UCSC release mm9) [ 43] using REad ALigner (REAL) [ 44, 45].
For differential expression analyses of MAP4 isoforms, the mouse genome assembly (release date July 2007 (NCBI37/mm9)) was examined using the UCSC Genome Browser at http://genome.ucsc.edu/.ucsc.edu/
We matched tags on the mouse genome (Ensembl release 47, based on NCBI m37 assembly) by using the Megablast algorithm [ 24].
The abundance of each signature was converted to transcripts per million (tpm), and the MPSS signatures were mapped to UniGene clusters by Lynx Therapeutics, based on the mouse genome sequence (Release #3, Feb 2003) [ 45] and the mouse UniGene sequences (UniGene Build #122) [ 46].
We used the mouse druggable genome release 1 and a customized transcription factor siRNA library (QIAGEN) designed against 8,296 and 1,640 genes, respectively.
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