treatment Protocols

Category: Nucleic Acid Electrophoresis Protocols: Pulsed Field Gel Electrophoresis Protocols

Protocol for retrieval of DNA fragments from pulsed-field gels following DNA concentration. DNA contained in a slice of low-melting-temperature agarose is first concentrated by electrophoresis into a high-percentage agarose gel, and then isolated by treatment with agarase.  The resulting DNA preparation is purified by microdialysis. - [Read Retrieval of DNA Fragments from Pulsed-field Gels following DNA Concentration Protocol]

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols: DNA Staining Protocols for Flow Cytometry

In an attempt to accurately measure DNA content with simultaneous preservation of cell surface markers, we have utilized gentle ethanol treatment techniques, which permeablize cells with minimal loss of surface antigen expression and antibody-antigen association. For some cell types, the presence of apoptotic cells based on reduced DNA content can also be detected. One such technique employs the addition of ethanol to cells previously resuspended in high concentrations of fetal bovine serum... - [Read Simultaneous Analysis of DNA Content and Surface Immunophenotype Protocol]

Category: Yeast Protocols: Yeast Nucleic Acid Protocols: Yeast PCR Protocols

Protocol for single tube confirmation yeast PCR. Includes: Clonal purification; Zymolyase treatment; PCR reactions; PCR conditions; Agarose gel electrophoresis. - [Read Single Tube Confirmation Yeast PCR Protocol]

Category: Genetics and Genomics: Genotyping Protocols

SNP Genotyping System. Includes: Protocol for Manual READIT® Assay in Multiwell Plates; Preparation of L/L Reagent; Preparation of Heating Block; CIAP/Exo Treatment; Preparation of Interrogation Probe Solutions; Template Denaturation; Sample Interrogation; Protocol for Manual Plate Reading Luminometers; Protocol for Injecting Plate Reading Luminometers; Protocol for Automated READIT® Assay with the Biomek® 2000 Instrument. - [Read SNP Genotyping System]

Category: Nucleic Acid Electrophoresis Protocols: RNA Electrophoresis Protocols

Protocol describes the transfer of RNA from agarose gels to neutral or positively charged nylon membranes, using upward capillary flow of neutral or alkaline buffers.  RNA becomes covalently fixed to positively charged nylon membranes during transfer in alkaline buffers.  However, treatment by UV irradiation or heating is required to fix RNA to neutral membranes. - [Read Transfer and Fixation of Denatured RNA to Membranes Protocol]

Category: Epigenetics and Methylation Protocols: 5-Aza-dC Treatment AZA Procedure

"Cells were seeded at a density of 1X106 cells/100-mm dish with 10% FBS and allowed to attach over a 24-h period. 5-Aza-dC (Sigma) was then added to a final concentration of 0.2–1
M, and the cells were allowed to grow for 5 days. The medium with or w - [Read Treatment of Cells with 5-aza-dC. protocol PDF]

Category: Drosophila Protocols: Drosophila Genetics Protocols

Protocol for whole mount in situ hybridization for the detection of mRNA in Drosophila embryos. Includes: Embryo collection and fixation; Prehybridization treatment; Hybridization; Washing and detection. - [Read Whole Mount In Situ Hybridization for the Detection of mRNA in Drosophila Embryos Protocol]