Protocol describes typical methods that are used to propagate and purify AAV vectors for experiments both in vitro and in vivo. Includes: Principles of the Triple Plasmid Transfection System; Plasmids; Transfection and Extraction of Virus; Purification of the AAV vector. - [Read A Protocol for AAV Vector Production and Purification]
Protocol is used to propagate and purify AAV vectors for experiments both in vitro and in vivo. Includes: Principles of the Triple Plasmid Transfection System; Plasmids; Transfection and Extraction of Virus; Purification of the AAV vector. - [Read AAV Vector Production and Purification Protocol]
Anatomy of a comparative gene expression study. Includes: Choosing Cell Populations; mRNA Extraction and Reverse Transcription; Fluorescent Labeling of cDNA's; Hybridization to a DNA Microarray; Scanning the Hybridized Array; Interpreting the Scanned Image. - [Read Anatomy of a Comparative Gene Expression Study]
Information on anatomy of a comparative gene expression study. Includes: Choosing cell populations; mRNA Extraction and Reverse Transcription; Fluorescent labeling of cDNA's; Hybridization of DNA microarray; Scanning the hybridized array; Interpreting the scanned image. - [Read Anatomy of a Comparative Gene Expression Study]
A protocol for extraction or isolation of both DNA and RNA from the same material, typically plant leaf or leaves. - [Read DNA and RNA Extraction Protocols]
DNA Extraction from Agarose Gels Protocol. The page includes cutting out the DNA band from the gel, and describes three methods including 1) Spin-columns (Nucleic acid purification columns), 2) using Dialysis tubing (semi-permeable membrane, Visking tubing), and the 3) Paper strip method.Matt Lewis, Department of Pathology, University of Liverpool. - [Read DNA Extraction from Agarose Gels Protocol]
Purifying nuclear pellets, Extraction of DNA from nuclei. Method for DNA isolation from Blood. Genomic Variation Laboratory, UC Davis. - [Read DNA extraction from blood]
DNA extracted from peripheral blood leucocytes using 3mls of whole blood. Modification of a salting out procedure as described by Miller et al., (1988). The DNA Laboratory, Medical School, Malta. - [Read DNA Extraction From Blood Leukocytes]
Cheek cells obtained by rinsing the mouth commercial mouth wash solution. Mouth wash is then discarded into a sterile conical tube and sent to the lab. Based on Salting out procedure. DNA Laboratory, Medical School, Malta. - [Read DNA Extraction from Cheek Cells]
Protocol is the first in a set of three describing fluorescent mRNA differential display (FDD or FDDRT-PCR). The method begins with the harvesting of total RNA from the tissue-cultured cells of interest. For other starting materials, such as blood samples, please see Extraction and Purification of RNA from Blood Samples for Fluorescent mRNA Differential Display. - [Read Extraction and Purification of RNA from Tissue-Cultured Cells for Fluorescent mRNA Differential]
Protocol for extraction and purification of total RNA using TRIzol OR TRI reagent. Includes: Homogenization for Cell Suspensions; Phase Separation; RNA Precipitation; RNA Wash; Redissolving the RNA; Determination of RNA Concentration and Purity; Preparation of Rnase-free water. - [Read Extraction and Purification of Total RNA using TRIzol OR TRI Reagent Protocol]
The growth conditions of microbial cell cultures and the time of sample collection should be optimized and standardized when growing cells for protein extraction. Because cells may excrete proteases and other extracellular enzymes, and compounds in the medium may interfere with extraction, wash cultures with an isotonic buffer, such as PBS or sucrose before solubilization. - [Read Extraction and Solubilization of Total Protein from Microorganisms Protocol]
DNA microarrays are an ordered arrangement of DNA molecules complementary to genes of interest that are "spotted" by robotic equipment onto a glass slide substrate. The expression of genes in cells can be monitored with microarrays by preparing cDNA from the mRNA of cells of interest and measuring the hybridization to the microarray. This protocol describes the labeling of genomic DNA for use as a probe for hybridization to the cDNA spotted on the array.
A single step RNA isolation protocol using Phenol Chloroform Extraction and Acid Guanidinium Thiocyanate. This RNA isolation method uses the fact that guanidinium thiocyanate can simultaneously lyse the cells and inactive cellular RNAses during the initial RNA isolation step allow a single step in the method.