68 Result(s)
within
Protocol
**-** Ren
Life Sciences
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Protocol
Single-Cell Imaging of mRNA by Target RNA-Initiated RCA
We present a powerful method for direct mRNA detection based on ligation-based recognition and in situ amplification, capable of single-cell imaging mRNA at single-nucleotide and single-molecule resolution. At...
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Repurposing Decellularized Lung to Generate Vascularized Fat
Conventional therapies to address critically sized defects in subcutaneous adipose tissue remain a reconstructive challenge for surgeons, largely due to the lack of graft pre-vascularization. Adipose tissue re...
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Simple N-Glycan Profile Analysis Using Lectin Staining, Mass Spectrometry, and GlycoMaple
The Golgi apparatus is one of the major sites of protein and lipid glycosylation and processing. Protein N-glycosylation plays critical roles in protein folding, transport, stability, and activity. Various gly...
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Actin: Static and Dynamic Studies
The actin cytoskeleton is a highly dynamic network in plant cells, which is precisely regulated by numerous actin-binding proteins. Hence, characterizing the biochemical activities of actin-binding proteins is...
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Staining Starch with Iodine Solution
Starch is important material in plant tissues, especially for storage tissues. Starches from different plant resources or tissues vary in morphology, content, and physicochemical properties. Starch and iodine ...
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Methods to Visualize and Quantify Cortical Microtubule Arrays in Arabidopsis Conical Cells
Many studies from different model organisms have demonstrated that microtubules are essential for various cellular processes, including cell division, cell morphogenesis, and intracellular trafficking. In inte...
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Protoplast Isolation for Plant Single-Cell RNA-seq
The growth and development of plants depends on diversified gene expression in different cell types. Compared to traditional bulk RNA sequencing, droplet-based single-cell RNA sequencing (scRNA-seq) allows for...
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High-Throughput Screening for Thermostable Polyester Hydrolases
Due to the promise of more sustainable recycling of plastics through biocatalytic degradation, the search for and engineering of polyester hydrolases have become a thriving field of research. Furthermore, amon...
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Methylome Imputation by Methylation Patterns
DNA methylation is studied extensively for its relations with several biological processes such as transcriptional regulation. While methylation levels are usually estimated per cytosine or genomic region, add...
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Quantifying the Binding and Target-Search Kinetics of Transcriptional Regulatory Factors by Live-Cell Single-Molecule Tracking
Eukaryotic transcriptional regulatory factors, such as transcription factors and epigenetic regulatory factors, must locate, bind, and assemble at specific genomic regions to execute functions within the compl...
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Synthetic Biology Meets Machine Learning
This chapter outlines the myriad applications of machine learning (ML) in synthetic biology, specifically in engineering cell and protein activity, and metabolic pathways. Though by no means comprehensive, the...
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PAM-Less CRISPR-SpRY Genome Editing in Plants
Engineered SpCas9 variant, SpRY, has been demonstrated to facilitate protospacer adjacent motif (PAM) unrestricted targeting of genomic DNA in various biological systems. Here we describe fast, efficient, and ...
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Single-Cell Joint Profiling of Open Chromatin and Transcriptome by Paired-Seq
Simultaneous detection of chromatin accessibility and transcription from the same cells promises to greatly facilitate the dissection of cell-type-specific gene regulatory programs in complex tissues. Paired-s...
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Tissue Cultivation, Preparation, and Extraction of High Molecular Weight DNA for Single-Molecule Genome Sequencing of Plant-Associated Fungi
Extraction of high-quality, high molecular weight DNA is a critical step for sequencing an organism’s genome. For fungi, DNA extraction is often complicated by co-precipitation of secondary metabolites, the mo...
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Establishment of Embryonic Zebrafish Xenograft Assays to Investigate TGF-β Family Signaling in Human Breast Cancer Progression
Transforming growth factor-β (TGF-β) family members have pivotal functions in controlling breast cancer progression, acting not only on cancer cells but also on other cells within the tumor microenvironment. H...
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Real-Time Quantification of Cell Internalization Kinetics by Bioluminescent Probes
Alongside the intracellular transport of nutrients needed for cellular homeostasis, great efforts exist to effectively deliver substances such as proteins and genes into the cell for therapy, gene editing, dis...
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In Situ Imaging of mRNA Splicing Variants by SpliceRCA
We present a method for multiplexed in situ imaging of individual RNA splicing variants. This method enables quantifying RNA splicing variants with single-molecule resolution, discriminating splicing isoforms ...
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Methods to Investigate Cell Polarity of Inner Ear
Hair cells in cochlea and vestibular organs depend on the coordinated cell polarity to perform the normal auditory or balance function. The mouse inner ear is one of the ideal model to study planar cell polari...
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Nanobodies: From Serendipitous Discovery of Heavy Chain-Only Antibodies in Camelids to a Wide Range of Useful Applications
The presence of unique heavy chain-only antibodies (HCAbs) in camelids was discovered at Vrije Universiteit Brussel (VUB, Brussels, Belgium) at a time when many researchers were exploring the cloning and expre...
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Cytoplasmic Expression of Nanobodies with Formylglycine Generating Enzyme Tag and Conversion to a Bio-Orthogonal Aldehyde Group
Nanobodies (Nbs) can be successfully retrieved following phage, bacterial, yeast, or ribosome display of immune, synthetic, or naïve libraries. However, after panning, multiple individual Nb clones need to be ...
