skeletal muscle cdna library  (TaKaRa)

Journal: PLoS ONE

Article Title: Multiple Novel Nesprin-1 and Nesprin-2 Variants Act as Versatile Tissue-Specific Intracellular Scaffolds

doi: 10.1371/journal.pone.0040098

Figure Lengend Snippet: A) cDNA ends identified by 3′ and 5′ RACE from Brain, Skeletal Muscle (SkeMus) and HeLa cDNA libraries. B) DNA sequencing results suggest that nesprin isoforms terminate with unique C-terminal ends absent from the giant isoforms as a result of intron retention. For example, isoforms utilising the N1-3′E90 UTR terminate with ‘AGAGYPHQ’ amino acids, giving it a unique fingerprint. Blue sequences show the coding regions of exons 90 and 91, black sequences show intronic regions and red sequence indicates a stop codon. C) Validation and tissue specificity of nesprin-1 UTRs identified on online databases and by RACE were confirmed by PCR amplification from a multiple tissue cDNA panel and DNA sequencing. Nesprin-1 PCRs were carried out when UTRs were identified on cDNA panels available at the time and are therefore organised into 3 separate sections. D) Validation and tissue specificity of nesprin-2 UTRs identified on online databases and by RACE were confirmed by PCR amplification from a multiple tissue cDNA panel and DNA sequencing. Nesprin-2 PCRs were carried out when UTRs were identified on cDNA panels available at the time and are therefore organised into 3 separate sections. Small Intestine and Peripheral Blood Lymphocytes have been abbreviated as ‘SI’ and ‘PBL’ respectively for all cDNA panels.

Article Snippet: Rapid Amplification of cDNA Ends (RACE) on Brain, HeLa and Skeletal muscle Marathon-Ready cDNA libraries using the Advantage-2 PCR kit (Clontech) and gene specific primers was performed ( ).

Techniques: DNA Sequencing, Sequencing, Amplification

Journal: PLoS ONE

Article Title: Multiple Novel Nesprin-1 and Nesprin-2 Variants Act as Versatile Tissue-Specific Intracellular Scaffolds

doi: 10.1371/journal.pone.0040098

Figure Lengend Snippet: A) PCR amplification across splice sites was carried out from cDNA isolated from U2OS cells. Splicing of exon 93 for nesprin-1 was observed as was the splicing for nesprin-2 exon 107’. B) PCR amplification across splice sites was carried out from cDNA isolated from VSMCs. Splicing of exon 93 for nesprin-1 was observed. Exon 107’ was retained in all nesprin-2 transcripts while splicing of exons 110–113 was also observed in these cells. + Represents bands with exon(s) excluded.

Article Snippet: Rapid Amplification of cDNA Ends (RACE) on Brain, HeLa and Skeletal muscle Marathon-Ready cDNA libraries using the Advantage-2 PCR kit (Clontech) and gene specific primers was performed ( ).

Techniques: Amplification, Isolation

Journal: International Journal of Biological Sciences

Article Title: Direct Evidence for Calcineurin Binding to the Exon-7 Loop of the Sodium-Bicarbonate Cotransporter NBCn1

doi: 10.7150/ijbs.9539

Figure Lengend Snippet: NBCn1-Exon 7 amplified from human tissues. A. Nested primers designed from the human genome upstream of the MERF initiation site were used in PCR amplification of cDNA from skeletal muscle (lane 2) and kidney (lane 4) libraries. Products were separated on a 1% agarose gel. The products are ~3.7 kbp, as judged by comparison with the 1-kb ladder (lane 1). After sequencing, the bands were identified as genes encoding NBCn1 containing Exon 7. The isolated kidney clone is identical to that of the NBC3 clone reported by the Kurtz group , , except for various point mutations and polymorphisms. No clones were amplified from liver (lane 3) with these particular primers. B. Examples of NBCn1 clones with a MEAD initiation site are shown as amplified in skeletal muscle. Two bands (~3.4 kbp and ~3.7 kbp; lane 2) encoding NBCn1 were observed, as judged by comparison with the 2-log DNA marker (lane 1). The upper band encodes for NBCn1 with Exon 7, and the lower is without Exon 7. Similarly, an NBCn1-Exon 7 clone was amplified from liver (lane 4), and detected by internal gene-specific primers corresponding to an expected ~2.8-kbp product. C. A summary of the tissue distribution found for the Nt of NBCn1 clones (with and without Exon 7) is shown in all experiments. There appears to be a complementary distribution of expression between Nt-Exon 7 clones that begin with MEAD versus MERF. Large (easily detectable) and small (barely detectable) " + " signs indicate relative amplification intensity of signals using our primers.

Article Snippet: NBCn1 splice variants were amplified by nested PCR reactions using human cDNA libraries, having adaptor-ligated AP1-ends, from kidney, skeletal muscle, and liver tissues (Clontech, CA).

Techniques: Amplification, Agarose Gel Electrophoresis, Sequencing, Isolation, Clone Assay, Marker, Expressing

Journal: eLife

Article Title: Myopalladin knockout mice develop cardiac dilation and show a maladaptive response to mechanical pressure overload

doi: 10.7554/eLife.58313

Figure Lengend Snippet:

Article Snippet: Recombinant DNA reagent , Human skeletal muscle cDNA library in the pGAD10 vector , Clontech Laboratories , HL4010AB , .

Techniques: Knock-Out, Recombinant, Modification, Plasmid Preparation, cDNA Library Assay, Clone Assay, Sequencing, Labeling, DC Protein Assay, Transformation Assay, Protease Inhibitor, Western Blot, Staining, Software