Average gene expression stability values of the candidate reference genes in the derived and produced embryo samples as calculated by geNorm software and ranking made based on the relative stability values

<p><b>Copyright information:</b></p><p>Taken from "Expression profiles of the pluripotency marker gene and validation of reference genes in rabbit oocytes and preimplantation stage embryos"</p><p>http://www.biomedcentral.com/1471-2199/9/67</p><p>BMC Molecular Biology 2008;9():67-67.</p><p>Published online 28 Jul 2008</p><p>PMCID:PMC2507718.</p><p></p

Mitochondrial DNA heteroplasmy in ovine fetuses and sheep cloned by somatic cell nuclear transfer-3

-oocyte mtDNA) and ARMS-qPCR (non-discriminative (ND) and discriminative (D) assays) for the low-abundant SNP variant (target here: donor mtDNA) at the mutated site. The amplification efficiency of the D assay was 91%. Without further optimization concerning conditions for enzymatic digestion, quantification by REMS-qPCR targeting a single donor-B-specific SNP reached a detection limit of 0.02%, i.e. a point mutation discrimination selectivity factor of 5 × 10. It allowed detection of heteroplasmy (0.1%) in the donor B-derived clone CB2. This could not be detected by conventional ARMS-qPCR discriminating point mutations only down to 0.1% (see: illustrated improvement of discrimination). For clarity each plot is presented as the mean calculated from duplicate amplification reactions. Individual Ct values, i.e. PCR cycle numbers at which plots crossed an arbitrarily placed signal threshold, are given in the figure key. An independent technical replicate of this REMS-qPCR experiment demonstrated reproducibility of the method (data not shown).<p><b>Copyright information:</b></p><p>Taken from "Mitochondrial DNA heteroplasmy in ovine fetuses and sheep cloned by somatic cell nuclear transfer"</p><p>http://www.biomedcentral.com/1471-213X/7/141</p><p>BMC Developmental Biology 2007;7():141-141.</p><p>Published online 21 Dec 2007</p><p>PMCID:PMC2323970.</p><p></p

Mitochondrial DNA heteroplasmy in ovine fetuses and sheep cloned by somatic cell nuclear transfer-1

-oocyte mtDNA) and ARMS-qPCR (non-discriminative (ND) and discriminative (D) assays) for the low-abundant SNP variant (target here: donor mtDNA) at the mutated site. The amplification efficiency of the D assay was 91%. Without further optimization concerning conditions for enzymatic digestion, quantification by REMS-qPCR targeting a single donor-B-specific SNP reached a detection limit of 0.02%, i.e. a point mutation discrimination selectivity factor of 5 × 10. It allowed detection of heteroplasmy (0.1%) in the donor B-derived clone CB2. This could not be detected by conventional ARMS-qPCR discriminating point mutations only down to 0.1% (see: illustrated improvement of discrimination). For clarity each plot is presented as the mean calculated from duplicate amplification reactions. Individual Ct values, i.e. PCR cycle numbers at which plots crossed an arbitrarily placed signal threshold, are given in the figure key. An independent technical replicate of this REMS-qPCR experiment demonstrated reproducibility of the method (data not shown).<p><b>Copyright information:</b></p><p>Taken from "Mitochondrial DNA heteroplasmy in ovine fetuses and sheep cloned by somatic cell nuclear transfer"</p><p>http://www.biomedcentral.com/1471-213X/7/141</p><p>BMC Developmental Biology 2007;7():141-141.</p><p>Published online 21 Dec 2007</p><p>PMCID:PMC2323970.</p><p></p

Mitochondrial DNA heteroplasmy in ovine fetuses and sheep cloned by somatic cell nuclear transfer-2

Tings from an alignment of concatenated mtDNA sequences (3806 nucleotides: control region, , , and ) of the three donor cell lines, their SCNT clones (CA1 to CA7, CB1 to CB3, and CC1 and CC2) and of an European mitochondrial reference genome (GenBank:). The scale bar represents 0.001 substitutions per site, and quartet puzzling values are shown (all are >50). The numbers at the nodes (quartet puzzling values) indicate the frequencies of occurrence for 1,000 replicate trees. Quartet puzzling support values provide an estimate of support of a given branch and can be interpreted in much the same way as bootstrap values. CA5 is the most divergent donor A-derived clone which is highlighted by the ellipse (see below for amino acid changes).<p><b>Copyright information:</b></p><p>Taken from "Mitochondrial DNA heteroplasmy in ovine fetuses and sheep cloned by somatic cell nuclear transfer"</p><p>http://www.biomedcentral.com/1471-213X/7/141</p><p>BMC Developmental Biology 2007;7():141-141.</p><p>Published online 21 Dec 2007</p><p>PMCID:PMC2323970.</p><p></p

Mitochondrial DNA heteroplasmy in ovine fetuses and sheep cloned by somatic cell nuclear transfer-0

Tings from an alignment of concatenated mtDNA sequences (3806 nucleotides: control region, , , and ) of the three donor cell lines, their SCNT clones (CA1 to CA7, CB1 to CB3, and CC1 and CC2) and of an European mitochondrial reference genome (GenBank:). The scale bar represents 0.001 substitutions per site, and quartet puzzling values are shown (all are >50). The numbers at the nodes (quartet puzzling values) indicate the frequencies of occurrence for 1,000 replicate trees. Quartet puzzling support values provide an estimate of support of a given branch and can be interpreted in much the same way as bootstrap values. CA5 is the most divergent donor A-derived clone which is highlighted by the ellipse (see below for amino acid changes).<p><b>Copyright information:</b></p><p>Taken from "Mitochondrial DNA heteroplasmy in ovine fetuses and sheep cloned by somatic cell nuclear transfer"</p><p>http://www.biomedcentral.com/1471-213X/7/141</p><p>BMC Developmental Biology 2007;7():141-141.</p><p>Published online 21 Dec 2007</p><p>PMCID:PMC2323970.</p><p></p