cycle Protocols

Category: PCR Protocols: Inverse PCR Protocols

Protocol for inverse PCR & cycle sequencing of P element insertions for STS generation. - [Read Inverse PCR & Cycle Sequencing of P Element Insertions for STS Generation Protocol]

Category: Tetrahymena Protozoa Protocols

Protocol describes how to allow the isolation of nuclei from all stages of the Tetrahymena life cycle in high yield with a high degree of purity. This method gives highly purified populations of both micronuclei and macronuclei. - [Read Isolation and Purification of Tetrahymena Nuclei Protocol]

Category: Developmental Biology

Describe the use of replication-incompetent retroviral vectors for the analysis of lineal relationships in developing vertebrate tissues. An overview of the relevant aspects of the retroviral life cycle, and the strategies and current methods in use at their laboratory are described. - [Read Lineage Analysis Using Retroviral Vectors]

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols: Cell Cycle Analysis Protocols

Exponentially growing cells are asynchronous with respect to the cell cycle stage. Detection of cell cycle-related events is improved by enriching the culture for cells at the stage during which the particular event occurs. Methods for synchronizing cells are provided here, including those based on morphological features of the cell. - [Read Methods for Synchronizing Cells at Specific Stages of the Cell Cycle]

Category: RNA Protocols: cDNA Libraries Library

(by PCR of phage insert, cycle sequencing using ABI dye terminator kits and automated sequencing on an ABI 373A) Ian Ridgers - [Read Natural History Museum Protocol for EST sequencing from lambda zap phage plaques]

Category: DNA Protocols: DNA Sequencing Protocols: Big Dye Protocols

Protocol describes the purification, quantification, and subsequent sequencing of amplified DNA fragments using PCR. Excess nucleotides are removed from the initial PCR products using spun columns, and the products are quantified using fluorometry. - [Read Nonradioactive Cycle Sequencing of PCR-Amplified DNA Protocol]

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols: Flow Cytometry Immunofluorescence Staining Protocols

Protocol for phenotype-specific immunodetection of cyclins using 488/630 nm dual laser flow cytometry. This protocol is for use with the D and E cyclins and employs 488 nm argon laser excitation of propidium iodide and a FITC-conjugated phenotypic label, and 630 nm NeNe or diode laser excitation of the fluorochrome Cy5 to detect cell cycle-specific cyclin D expression. - [Read Phenotype-Specific Immunodetection of Cyclins using 488/630 nm Dual Laser Flow Cytometry Protocol]

Category: Yeast Protocols: Yeast Nucleic Acid Protocols

Flow cytometry is used to analyze the quantity of DNA in cells. Since the DNA content of cells varies through the cell cycle, this information can provide an indication of cell cycle progression. This protocol uses SYTOX Green staining. - [Read Preparation of Yeast Cells for Flow Cytometry Protocol]

Category: Cell Biology and Cell Culture: Cell Cycle Protocols

This method works well to assess cell cycle distribution of whole cell populations. This method can also be used to assess the cell cycle distribution of GFP transfected cells however, the EtOH step is generally not sufficient to keep GFP in the cell. - [Read Preparing Cells for PI/FACS (cell cycle) Analysis Protocol]

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols: Cell Cycle Analysis Protocols

Protocol for preparing cells for PI/FACS (cell cycle) analysis. - [Read Preparing Cells for PI/FACS (cell cycle) Analysis Protocol]

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols: Flow Cytometry Data Analysis Protocols

Protocol for preparing cells for PI/FACS (cell cycle) analysis. - [Read Preparing Cells for PI/FACS (cell cycle) Analysis Protocol]

Category: Immunology: B Cell Protocols

The water soluble, DNA intercalator, propidium iodide (PI), is used to bind to DNA after permeabilization of cells with NP40. The amount of dye bound correlates with the content of DNA within a given cell. Once cells are stained, they are analyzed on a flow cytometer. The relative content of DNA indicates the distribution of a population of cells throughout the cell cycle. - [Read Quantification of Apoptosis and Cell Cycle Distribution of Primary B Cells Using Propidium Iodide]

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols: Apoptosis Cytometry Protocols

The water soluble, DNA intercalator, propidium iodide (PI), is used to bind to DNA after permeabilization of cells with NP40. The amount of dye bound correlates with the content of DNA within a given cell. Once cells are stained, they are analyzed on a flow cytometer. The relative content of DNA indicates the distribution of a population of cells throughout the cell cycle. - [Read Quantification of Apoptosis and the Cell Cycle Distribution of Primary B Cells Using PI]

Category: DNA Protocols: PCR Polymerase Chain Reaction Protocols: Long PCR Protocols

Long PCR Buffer, Cycle times and temperatures, Picking Primers, Hot Start Polymerases for Long PCR. Harvard-Lipper Center for Computational Genetics. Modified from Cheng et al. - [Read Reagents and Guidelines for Long PCR]

Category: PCR Protocols: Reverse Transcriptase RT-PCR Protocols

In the first part of this protocol, the linear range of amplification is determined by carrying out 10 identical PCRs in the presence of [{alpha}-32P]dCTP and stopping one reaction after every two cycles.  Amplification products are quantified on a denaturing polyacrylamide gel and the results plotted on a graph (counts per minute vs. cycle number).  Total RNA is used as an internal control. - [Read Relative RT-PCR: Determining the Linear Range of Amplification and Optimizing the Primers:Competimer]

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols: Apoptosis Cytometry Protocols

Two distinct modes of cell death, apoptosis and necrosis, can be distinguished on the basis of differences in morphological, biochemical, and molecular changes occurring in the dying cells. Report the development of flow cytometric techniques that permit concomitant detection of apoptosis and cellular DNA content or BrdU content analysis. - [Read The Use of Flow Cytometry for Concomitant Detection of Apoptosis and Cell Cycle Analysis]

Category: Cell Biology and Cell Culture: Cell Cycle Protocols

Protocol for yeast cell cycle by flow cytometry. Includes procedure for dyes: Propidium Iodide and Sytox Green. - [Read Yeast Cell Cycle by Flow Cytometry Protocol]

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols: Cell Cycle Analysis Protocols

Protocol for yeast cell cycle by flow cytometry. - [Read Yeast Cell Cycle by Flow Cytometry Protocol]

Category: Yeast Protocols

Protocol for Yeast cell cycle by flow cytometry. - [Read Yeast Cell Cycle by Flow Cytometry Protocol]