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IJMS | Free Full-Text | Myogenetic Oligodeoxynucleotide Induces Myocardial Differentiation of Murine Pluripotent Stem Cells

An 18-base myogenetic oligodeoxynucleotide (myoDN), iSN04, acts as an anti-nucleolin aptamer and induces myogenic differentiation of skeletal muscle myoblasts. This study investigated the effect of iSN04 on murine embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs). In the undifferentiated state, iSN04 inhibited the proliferation of ESCs and iPSCs but did not affect the expression of pluripotent markers. In the differentiating condition, iSN04 treatment of ESCs/iPSCs from day 5 onward dramatically induced differentiation into Nkx2-5+ beating cardiomyocytes with upregulation of Gata4, Isl1, and Nkx2-5, whereas iSN04 treatment from earlier stages completely inhibited cardiomyogenesis. RNA sequencing revealed that iSN04 treatment from day 5 onward contributes to the generation of cardiac progenitors by modulating the Wnt signaling pathway. Immunostaining showed that iSN04 suppressed the cytoplasmic translocation of nucleolin and restricted it to the nucleoli. These resu ....

United Kingdom , United States , Baden Wüberg , San Diego , Jackson Immunoresearch , Wayne Rasband , National Bio , National Institute Of Health , Myocardial Differentiation , Mesoderm Differentiation , Wnt Signaling Pathway , Nucleolin Translocation , Not Affect Myocardial Cell , Fujifilm Wako Chemicals , Regenerative Medicine , National Bio Resource Project , Thermo Fisher Scientific , National Institute , West Grove , Revertra Ace , Stepone Real Time , Agilent Technologies , Illumina Truseq , Prep Kits , Life Technologies , Illumina Novaseq ,

mRNA vaccine quality analysis using RNA sequencing | Nature Communications

The success of mRNA vaccines has been realised, in part, by advances in manufacturing that enabled billions of doses to be produced at sufficient quality and safety. However, mRNA vaccines must be rigorously analysed to measure their integrity and detect contaminants that reduce their effectiveness and induce side-effects. Currently, mRNA vaccines and therapies are analysed using a range of time-consuming and costly methods. Here we describe a streamlined method to analyse mRNA vaccines and therapies using long-read nanopore sequencing. Compared to other industry-standard techniques, VAX-seq can comprehensively measure key mRNA vaccine quality attributes, including sequence, length, integrity, and purity. We also show how direct RNA sequencing can analyse mRNA chemistry, including the detection of nucleoside modifications. To support this approach, we provide supporting software to automatically report on mRNA and plasmid template quality and integrity. Given these advantages, we antic ....

Oxford Nanopore , Agilent Tapestation , Rna Stranded Library Preparation Kit , Illumina Truseq , Library Preparation Kit ,