Targeted disruption of the mouse gene encoding the cysteine- and glycine-rich LIM domain protein CRP2 result in subtle alteration of cardiac ultrastructure-0

M) controls by PCR to determine the chromosomal map location of . The mapping results of this analysis were deposited under accession no. MGI: 1202907 at the Jackson Laboratory and the murine gene localized to chromosome 10. In the figure, the entire T31 RH Chr 10 framework map is depicted on the left of the figure with the overall length calculated from the framework data []. The centromere is depicted by a black circle at the top of the map. The enlarged segment of distal Chromosome 10 is shown with respective framework markers listed to the left of the chromosome bar and a selection of mapped genes to the right. The distances between loci are calculated based on only the listed data sets, and unscored radiation hybrid cell line data are inferred where the data on either side of the missing score are in agreement. Blocks of human synteny are indicated to the right of the RH map, based on information from the NCBI's locus link []. Note all locus names should be in italics, but are shown in plain text for readability.<p><b>Copyright information:</b></p><p>Taken from "Targeted disruption of the mouse gene encoding the cysteine- and glycine-rich LIM domain protein CRP2 result in subtle alteration of cardiac ultrastructure"</p><p>http://www.biomedcentral.com/1471-213X/8/80</p><p>BMC Developmental Biology 2008;8():80-80.</p><p>Published online 19 Aug 2008</p><p>PMCID:PMC2529283.</p><p></p

Targeted disruption of the mouse gene encoding the cysteine- and glycine-rich LIM domain protein CRP2 result in subtle alteration of cardiac ultrastructure-3

In overlay in top two rows). Preimmune serum only picks up extracellular matrix in control sections (green signal in bottom two rows). The sections were counterstained for the Z-disc protein α-actinin (red signal in overlays) and with DAPI to visualise the nuclei (blue signal in overlays). The space bar represents 10 μM.<p><b>Copyright information:</b></p><p>Taken from "Targeted disruption of the mouse gene encoding the cysteine- and glycine-rich LIM domain protein CRP2 result in subtle alteration of cardiac ultrastructure"</p><p>http://www.biomedcentral.com/1471-213X/8/80</p><p>BMC Developmental Biology 2008;8():80-80.</p><p>Published online 19 Aug 2008</p><p>PMCID:PMC2529283.</p><p></p

Targeted disruption of the mouse gene encoding the cysteine- and glycine-rich LIM domain protein CRP2 result in subtle alteration of cardiac ultrastructure-2

Yte diameters using a microscope with an internal size scale. For details refer Material and Method section and Table 1. The space bar in each figure part represents 100 μM.<p><b>Copyright information:</b></p><p>Taken from "Targeted disruption of the mouse gene encoding the cysteine- and glycine-rich LIM domain protein CRP2 result in subtle alteration of cardiac ultrastructure"</p><p>http://www.biomedcentral.com/1471-213X/8/80</p><p>BMC Developmental Biology 2008;8():80-80.</p><p>Published online 19 Aug 2008</p><p>PMCID:PMC2529283.</p><p></p

Targeted disruption of the mouse gene encoding the cysteine- and glycine-rich LIM domain protein CRP2 result in subtle alteration of cardiac ultrastructure-6

M) controls by PCR to determine the chromosomal map location of . The mapping results of this analysis were deposited under accession no. MGI: 1202907 at the Jackson Laboratory and the murine gene localized to chromosome 10. In the figure, the entire T31 RH Chr 10 framework map is depicted on the left of the figure with the overall length calculated from the framework data []. The centromere is depicted by a black circle at the top of the map. The enlarged segment of distal Chromosome 10 is shown with respective framework markers listed to the left of the chromosome bar and a selection of mapped genes to the right. The distances between loci are calculated based on only the listed data sets, and unscored radiation hybrid cell line data are inferred where the data on either side of the missing score are in agreement. Blocks of human synteny are indicated to the right of the RH map, based on information from the NCBI's locus link []. Note all locus names should be in italics, but are shown in plain text for readability.<p><b>Copyright information:</b></p><p>Taken from "Targeted disruption of the mouse gene encoding the cysteine- and glycine-rich LIM domain protein CRP2 result in subtle alteration of cardiac ultrastructure"</p><p>http://www.biomedcentral.com/1471-213X/8/80</p><p>BMC Developmental Biology 2008;8():80-80.</p><p>Published online 19 Aug 2008</p><p>PMCID:PMC2529283.</p><p></p

Targeted disruption of the mouse gene encoding the cysteine- and glycine-rich LIM domain protein CRP2 result in subtle alteration of cardiac ultrastructure-5

Inal frozen sections of ventricular tissues from wild-type (panel ), (panel ) and (panel ) mice, stained with antibodies against β-catenin (panel ), connexin-43 (panel ) and N-RAP (panel ) antibodies. While β-catenin and N-RAP expression are significantly upregulated in deficient mice (panel B, H) and to a lesser extent in mice (panel C, I), connexin-43 expression is reduced in as well as mice (panel F). The space bar represents 10 μM.Western blot analysis of N-RAP and β-catenin. Heart extracts from normal (WT), - and MLP-deficient mice were probed with antibodies specific for N-RAP and β-catenin. Equal loading was demonstrated in Ponceau Red stain.<p><b>Copyright information:</b></p><p>Taken from "Targeted disruption of the mouse gene encoding the cysteine- and glycine-rich LIM domain protein CRP2 result in subtle alteration of cardiac ultrastructure"</p><p>http://www.biomedcentral.com/1471-213X/8/80</p><p>BMC Developmental Biology 2008;8():80-80.</p><p>Published online 19 Aug 2008</p><p>PMCID:PMC2529283.</p><p></p

Targeted disruption of the mouse gene encoding the cysteine- and glycine-rich LIM domain protein CRP2 result in subtle alteration of cardiac ultrastructure-1

G heterozygotes were discriminated by PCR analysis of tail biopsy DNAs. The positions of the oligonucleotide primers used for amplification of the wild-type (218 bp) and mutant (422 bp) PCR fragments are indicated. In the genotyping experiment shown, the different DNAs were genotyped as homozygous null (-/-), heterozygous (+/-) or wild-type (+/+). Southern hybridisation of littermate offspring from heterozygous intercrosses genotyped as heterozygote (WT), wild-type (WT), or homozygote null (). The DNA was digested with HI, fragments were separated in a 1% agarose gel and transferred to a nylon membrane. The blot was hybridized with an external 1.5 kbp I probe (see Methods), resulting in fragments of ~7.3 (knock out) or ~12.6 kb (wild-type) in size. Northern blot of total kidney RNA isolated from wild-type (+/+), heterozygote (+/-), and homozygote (-/-) null mice. The RNAs were hybridized with a specific cDNA probe. The autoradiograph showed the typical 1.2. kb signal in 2, a weaker band with , and a faint band with the mice. To verify the integrity of RNAs, the blot was subsequently hybridized with a -specific cDNA probe. Western blot of kidney homogenates extracted from wild-type (WT) and mutant mice (). As a positive control and to demonstrate the specificity of the CRP2 specific antibody, cell extracts taken from COS-7 cells that were transfected with myc-epitope tagged version of murine CRP1, CRP2 and CRP3 were taken. The expression of these proteins was demonstrated by subsequent probing with a myc-epitope specific antibody. In the nulls, no CRP2 band at any size was detected.<p><b>Copyright information:</b></p><p>Taken from "Targeted disruption of the mouse gene encoding the cysteine- and glycine-rich LIM domain protein CRP2 result in subtle alteration of cardiac ultrastructure"</p><p>http://www.biomedcentral.com/1471-213X/8/80</p><p>BMC Developmental Biology 2008;8():80-80.</p><p>Published online 19 Aug 2008</p><p>PMCID:PMC2529283.</p><p></p

Targeted disruption of the mouse gene encoding the cysteine- and glycine-rich LIM domain protein CRP2 result in subtle alteration of cardiac ultrastructure-4

He space bar represents 10 μM.Heart sections of wild-type mice (WT) and nulls () were infiltrated with an expoxy resin and examined in a Philips TEM 400 transmission microscope. The intercalated discs are each marked by arrowheads. Note the moderate and pronounced convolution of the membrane at the intercalated disc of null compared to wild type mice. (Original magnification × 9.000). Heart sections of wild-type mice (WT) and nulls () at higher magnification (× 18.000). For electron microscopic analysis three hearts taken from each genotype were analysed. The most representative images are shown in and .<p><b>Copyright information:</b></p><p>Taken from "Targeted disruption of the mouse gene encoding the cysteine- and glycine-rich LIM domain protein CRP2 result in subtle alteration of cardiac ultrastructure"</p><p>http://www.biomedcentral.com/1471-213X/8/80</p><p>BMC Developmental Biology 2008;8():80-80.</p><p>Published online 19 Aug 2008</p><p>PMCID:PMC2529283.</p><p></p

Comparison of five automated DNA extraction systems.

<p>Illustration of relative DNA concentrations of samples 1–10 measured by the NanoDrop 2000c spectrophotometer (A) and Qubit 2.0 fluorometer (B). The concentrations of the Maxwell 16 extracts were set to 100% for each sample.</p

Impact of DNA quantification methods on massively parallel sequencing.

<p>(A) Mean amplicon coverage determined by in-house software for each sample, quantification method and amount of DNA used for multiplex PCR amplification. (B) Number of all variants called by in-house software for each sample, quantification method and starting material used.</p

Impact of DNA quantification methods on downstream applications.

<p>(A) Electrophoretic pattern of 13 DNA extracts on a 1% agarose gel. (B) Amplified 275 bp fragment of the <i>EGFR</i> gene. 20 ng of sample DNA determined by each quantification method was used for PCR amplification. + indicates a positive control, - a negative control.</p