142 Result(s)
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Protocol
Purification of PCR Products from Agarose Gels for Direct Sequencing
The advent of direct sequencing of polymerase chain reaction (PCR) products has permitted extremely rapid analysis of DNA mutants and cDNA clones. However, direct PCR sequencing has been problematic for a numb...
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Protocol
Eukaryotic cell biosensor The Cytosensor microphysiometer
The Cytosensor, a silicon-based microphysiometer system, is a light-addressable potentiometric biosensor (LAPS) that detects functional responses of living cells in minutes. Thus, unlike other biosensors that ...
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Protocol
Preparation and Use of Thermosensitive Polymers
Restoration of organ structure and function, utilizing tissue engineering technologies, often requires the use of a temporary porous scaffold. The function of the scaffold is to direct the growth of cells migr...
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Protocol
Development of Female Contraceptive Vaccine Through DNA Inoculation of Human Chorionic Gonadotropin Beta Subunit (hCGß)
Human chorionic gonadotropin (hCG) has been considered as a primary target molecule for a contraceptive vaccine by the World Health Organization because of its physiological and temporal specificity. hCG is an...
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Protocol
Purification and Enzymatic Sequencing of Polymerase Chain Reaction Products
The advent of direct sequencing of polymerase chain reaction (PCR) products has permitted extremely rapid analysis of DNA mutants and cDNA clones. However, direct PCR sequencing has been problematic for a numb...
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Protocol
Smad Interactors in Bone Morphogenetic Protein Signaling
The yeast genetic-based two-hybrid system has been widely used in identifying a pair of interacting proteins. The basis for this technique is that eukaryotic transcription activators are modular with respect t...
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Protocol
Applying the Xenopus Oocyte Expression System to the Analysis of Gap Junction Proteins
The utility of Xenopus oocytes as a system for the expression and electrophysiological analysis of many ion channels has been well documented. This system offers several advantages. The oocytes are robust in term...
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Protocol
Use of Green Fluorescent Protein (GFP) to Study Cellular Dynamics
The green fluorescent protein (GFP) from the Pacific Northwest jellyfish Aequorea victoria is a member of a small but important class of proteins that exhibit strong visible fluorescence without the requirement o...
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Protocol
Introduction of NOS II Gene into Primary Cultures of Bovine and Human Endothelial Cells
In recent years, endothelial cells, activated by cytokines (1) have been found to phagocytose Gram-positive and Gram-negative bacteria (2–14), and exhibit microbicidal activity toward S. aureus (15). One of the e...
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Protocol
Labeling Stem Cells In Vitro for Identification of Their Differentiated Phenotypes After Grafting into the CNS
Stem cells have unlimited therapeutic potential for restoring CNS function lost secondary to trauma or degenerative disease. To date, stem cell grafts have only in a few instances partially ameliorated functio...
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Protocol
Direct Cloning of Full-Length Cell Differentially Expressed Genes by Multiple Rounds of Subtractive Hybridization Based on Long-Distance PCR and Magnetic Beads
Subtractive cloning is an ideal technique for identifying genes differentially expressed in two nuclear acids population (1). The polymerase chain reaction (PCR)-based subtraction is the method of choice when the...
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Protocol
in situ Immuno-PCR
Immunocytochemistry and in situ hybridization—the in situ staining approaches for detection of antigens, DNA, or RNA in intact cells and tissue sections— are common and powerful tools for biological and biomedica...
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Protocol
Construction of Genomic Libraries in λ-Vectors
Lambda (λ) bacteriophages are viruses that specifically infect bacteria. The genome of λ-phage is a double-stranded DNA molecule approx 50 kb in length (1). In bacterial cells, λ-phage employs one of two pathways...
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Protocol
Gene Transfer to Skeletal Muscle Using Herpes Simplex Virus-Based Vectors
Type 1 herpes simplex virus (HSV-1)-based vectors, which are naturally capable of carrying large DNA fragments like the 14 kb dystrophin cDNA, have been studied for their ability to transduce muscle cells (1–5). ...
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Protocol
Production and Surface Modification of Polylactide-Based Polymeric Scaffolds for Soft-Tissue Engineering
Current research in the field of tissue engineering is focused on the development of appropriate strategies for repair and regeneration of biological tissues. Biological tissues consist of cells situated withi...
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Protocol
Structural Determinants of Folate and Antifolate Membrane Transport by the Reduced Folate Carrier
Besides its important role in folate homeostasis, membrane transport is a critical determinant of the antitumor activities of antifolate therapeutics used in cancer chemotherapy, such as methotrexate (MTX) and an...
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Protocol
Differentiation of Human Embryonic Stem Cells Toward the Chondrogenic Lineage
Human embryonic stem cells (hESCs) have the ability to self-replicate and differentiate into cells from all three embryonic germ layers, thereby holding great promise for tissue regeneration applications. Howe...
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Protocol
Labeling Stem Cells In Vitro for Identification of their Differentiated Phenotypes after Grafting into the CNS
Grafting neural stem cells is a widely used experimental approach to central nervous system (CNS) repair after trauma or neurodegeneration. It is likely to be a realistic clinical therapy for human CNS disorde...
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Protocol
Molecular Beacons for Protein—DNA Interaction Studies
Real-time monitoring of DNA—protein interactions involving molecular beacon (MB) and molecular beacon aptamer (MBA) was discussed in this chapter. MBs are single-stranded oligonucleotide probes with a hairpin ...
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Protocol
Exocytosis of Endothelial Cells Is Regulated by N-Ethylmaleimide-Sensitive Factor
Endothelial exocytosis of granules is a rapid response to vascular injury. However, the molecular machinery that regulates exocytosis in endothelial cells is not well understood. Recently developed techniques ...
