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The causal mutation(s) for this QTL is expected to be located within this region of high homozygosity.

Like these previous methods, our method is just the first step to identify candidate genes or interaction pathways, enabling the search for causal mutation(s) for a phenotype whether SNP, indel or major mutation, to be narrowed.

From the initial 233 phenotypic series involving 1356 genes, we eventually tested 88 series that contained at least four genes and with known HGMD mutation(s) for the disease described in the phenotypic series, corresponding to 285 genes.

PCR-based one-step site-directed mutagenesis (SDM) is an efficient and rapid method to generate gene mutation(s) for study of protein structure-function relationship, identification of gene expression and modification of vector [ 1- 3].

Since then, we have mapped and identified the causative gene and mutation(s) for 12 and eight diseases, respectively, including congenital muscular dystonia I and II, crooked tail syndrome and stunted growth in BBCB (Charlier et al., 2008; Fasquelle et al., 2009; Sartelet et al., 2012a; Sartelet et al., 2012b).

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For the systematic identification of the mutation(s) responsible for the HL tolerant phenotype in hit1 and hit2, we applied a methodical workflow based on open source bioinformatics tools that allow quality filtering as well as sorting out common variants.

To map the amino acid mutation(s) responsible for acquisition of the cell cell fusion activity of S CK), three chimeric constructs were first made.

To identify the mutation(s) responsible for the unexpected resistance to TFV-DF we sequenced the entire RT coding region at time-points 1 and 2 (Figure S1, GenBank Accession Number AB506802 and AB506803).

Thus, the mutation(s) responsible for reduced fitness emerged later.

In the case of SBTD, such a change would have to be derived from a gene mutation(s) responsible for cryptorchidism.

Our recent discovery of the sexual cycle of S. rosetta (Levin and King, 2013) suggested that it might be possible to perform a choanoflagellate mapping cross to identify the mutation(s) responsible for the Rosetteless phenotype.

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