unique Protocols

Category: DNA Protocols: PCR Polymerase Chain Reaction Protocols: Standard PCR Protocol

"CORE SAMPLE" PCR - A method to re-PCR unique bands from products of mixed size - Molecular Biology Techniques Manual - Rybicki - [Read "CORE SAMPLE" PCR - A method to re-PCR unique bands from products of mixed size]

Category: PCR Protocols: Standard PCR Protocol

"CORE SAMPLE" PCR - A method to re-PCR unique bands from products of mixed size - Molecular Biology Techniques Manual - Rybicki - [Read "CORE SAMPLE" PCR - A method to re-PCR unique bands from products of mixed size]

Category: PCR Protocols: cDNA Synthesis Protocols

To generate "3'-end" partial cDNA clones, mRNA is reverse-transcribed using a "hybrid" primer (Qtotal, QT) that consists of two mixed bases (GATC/GAC followed by [T]17) and a unique 35-base oligonucleotide sequence (QI-QO).  Amplification is then performed using a primer containing part of this sequence (Qouter, Qo) (which now binds to each cDNA at its 3'-end) and a primer derived from the gene of interest, GSP1 (gene-specific primer 1). - [Read 3'-End cDNA Amplification Using Classic RACE Protocol]

Category: Cytogenetics Protocols: Fluorescent In Situ Hybridization FISH Protocol

Method assesses cellular mRNA transcripts in tissue sections and cell cultures using unique short anti-sense primers directed against sequences in particular protein(s). The unlabeled synthetic cDNA oligonucleotide primers are extended complementary to a sense mRNA transcript using reverse transcriptase and labeled through incorporation of a fluorescent-labeled dUTP nucleotide base. This procedure provides rapid detection of low abundance mRNA messages that can be related to other cellular.... - [Read Fluorescent In Situ Transcription in Cells and Tissues Protocol]

Category: Cell Organelle Protocols: Subcellular Fractionation Protocols

Acidocalcisomes, the dense acidic calcium-storing organelles, which were originally identified in Trypanosoma cruzi, have no parallels in mammalian cells. They thus represent a unique functional characteristic, not shared by the host and hence offer an important potential target for chemotherapy of Chagas disease. - [Read Fractionation of Acidocalcisomes and Other Organelles from Trypanosoma, Leishmania, Chlamydomonas]

Category: Yeast Protocols: Yeast Cell Synchrony Protocols

It is sometimes desirable to have an entire population of yeast cells that is growing synchronously or is arrested at a unique point in the cell cycle. - [Read Inducing Yeast Cell Synchrony: alpha-Factor Arrest Using bar1 Mutants Protocol]

Category: Yeast Protocols: Yeast Cell Synchrony Protocols

It is sometimes desirable to have an entire population of yeast cells that is growing synchronously or is arrested at a unique point in the cell cycle. - [Read Inducing Yeast Cell Synchrony: alpha-Factor Arrest Using Low Cell Concentration Protocol]

Category: Yeast Protocols: Yeast Cell Synchrony Protocols

It is sometimes desirable to have an entire population of yeast cells that is growing synchronously or is arrested at a unique point in the cell cycle. - [Read Inducing Yeast Cell Synchrony: alpha-Factor Arrest Using Low pH Protocol]

Category: Yeast Protocols: Yeast Cell Synchrony Protocols

It is sometimes desirable to have an entire population of yeast cells that is growing synchronously or is arrested at a unique point in the cell cycle. - [Read Inducing Yeast Cell Synchrony: Dilution of Stationary-Phase Cultures Protocol]

Category: Yeast Protocols: Yeast Cell Synchrony Protocols

It is sometimes desirable to have an entire population of yeast cells that is growing synchronously or is arrested at a unique point in the cell cycle. - [Read Inducing Yeast Cell Synchrony: Hydroxyurea Arrest Protocol]

Category: Yeast Protocols: Yeast Cell Synchrony Protocols

It is sometimes desirable to have an entire population of yeast cells that is growing synchronously or is arrested at a unique point in the cell cycle. - [Read Inducing Yeast Cell Synchrony: Nocodazole Arrest Protocol]

Category: Primary Cell Isolation and Culture Protocols: Laser Capture Microdissection Protocols

Human tissues are comprised of multiple interacting cell populations in a complex three dimensional arrangement with each cellular phenotype determined by a unique profile of mRNA and protein expression. Before microdissection techniques were developed, the only analysis tools for phenotypic studies were primarily immunohistochemistry and in-situ hybridization. While useful, these tools are limited to single gene analysis and, in general, do not allow qualitative studies. - [Read Microdissection Overview]

Category: Cloning Protocols: Mutagenesis Protocols

Protocol for oligonucleotide-directed mutagenesis by elimination of a unique restriction site (USE mutagenesis). - [Read Oligonucleotide-directed Mutagenesis by Elimination of a Unique Restriction Site (USE Mutagenesis)]

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols

Provides an overview of terminology and echniques unique to flow ytometry and is divided into two sections. The first section discusses technical aspects of flow cytometry which apply primarily to the instrumentation-oriented flow cytometry phase. The second section discusses electronic cell separation using flow cytometry. - [Read Overview of Flow Cytometry]

Category: Nucleic Acid Modification Protocols: DIG Biotin Labeled Probes Protocols

Protocol for PCR labeling of ds DNA with the PCR DIG Probe Synthesis Kit or PCR Labeling Mixes. Includes: PCR DIG labeling reaction for highly labeled probes containing unique sequences; PCR DIG labeling reaction for moderately labeled probes; PCR fluorescein labeling reaction for direct in situ probes. - [Read PCR Labeling of ds DNA with the PCR DIG Probe Synthesis Kit or PCR Labeling Mixes Protocol]

Category: Yeast Cell Culture Protocols: Yeast Sporulation Protocols

Zygotes can be identified by their unique morphology. They can be easily separated away from nonmated cells using a micromanipulator. This method provides an alternative to the selection of diploid cells on a medium that prevents the growth of haploid parent cells. - [Read Picking Zygotes Protocol]

Category: Cell Organelle Protocols: Subcellular Fractionation Protocols: Peroxisome Isolation Protocols

Peroxisomes can be purified in iodixanol gradients in high yield (80-90%) with no detectable contamination from any other organelle. This is a property unique to iodixanol because the densities of other organelles, particularly that of mitochondria (approx ρ = 1.14 g/ml) and endoplasmic reticulum (approx ρ = 1.13 g/ml) are much lower than that of peroxisomes (approx ρ = 1.18 g/ml). - [Read Purification of Peroxisomes using a Density Barrier in a Swinging-Bucket Rotor]

Category: Microscopy Protocols: In Vivo Imaging Protocols

Sophisticated fluorescence microscopy methods & equipment, now allow cellular events to be studied at high resolution in living material. The studying of living fly tissues presents unique difficulties in keeping the cells alive, introducing fluorescent probes, & imaging through thick hazy cytoplasm. This protocol outlines the preparation of major tissue types amenable to study by time-lapse cinematography and different methods for keeping them alive. - [Read Time-Lapse Cinematography in Living Drosophila Tissues: Preparation of Material]

Category: Genetics and Genomics: Genotyping Protocols: PCR Genotyping

This protocol is designed to detect sequences in the murine genome by polymerase chain reaction amplification, and is adapted from Stratman and Simon (Transgenic Res. 12, 521-522 (2003)).For those familiar with PCR genotyping, this method differs from the typical protocol by utilizing a unique enzyme (Klentaq), 30mer primers, and a 68° annealing temperature. - [Read Universal Mouse Genotyping Protocol]