TCA-DOC Normal, TCA Acetone Precipitation, Ethanol Precipitation, TCA-DOC / Acetone, Acidified Acetone/Methanol, Dr. Mario Lebendiker. University of Jerusalem. - [Read Protein Precipitation Protocols]
Protein Precipitation. Developed by: Eric Burtson. American Association of Immunologists 1995. In this lab you will separate a solution of proteins using protein precipitation. Since you will be testing the same protein mix that you used in the Gel Filtration lesson, you will pass the protein through a gel filtration column to identify which protein(s) precipitates. - [Read Protein Precipitation]
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]
A protein precipitation procedure used to precipitate protein from cell
lysates. Allows for optimal protein recovery and accurate assays. The Signaling Gateway - [Read TCA/Acetone Precipitation (Large Scale) PDF]
Agarose gel purification,
Annealing and extending, oligonucleotides, Ethanol Precipitation, Ligation
Miniprep, Oligonucleotide purification, Recovering DNA bands, Restriction digest
Gene Clean. The Hu Lab. - [Read DNA cloning Procedures]
Protein extraction from whole tissues for isoelectric focusing. University of Missouri - Columbia Proteomics Center.
SDS extraction followed by acetone precipitation. Also has Phenol extraction followed by methanolic ammonium acetate precipitation. - [Read Protein extraction from whole tissues for isoelectric focusing]
Methanol-Chloroform Precipitation of Proteins. A procedure for precipitating proteins from solution, including detergent solutions, is from Wessel and Flügge (1984). Univ. Virginia. - [Read Methanol-Chloroform Precipitation of Proteins]
Protocol is based on the standard Trizol protocol for the purification of RNA from animal cells using Trizol (Purification of RNA from Animal Cells using Trizol). In this version, adapted for use with plant tissues, a high-salt isopropanol precipitation step has been added to precipitate RNA selectively, while maintaining polysaccharides and proteoglycans in solution. - [Read Preparation of RNA from Plant Tissue Using Trizol]
Protocol uses the BIOPRIME reaction kit from GibcoBRL to prepare biotin-labelled BAC DNA which is detected using FITC-Avidin (Vector Labs, DCS grade). Reagents from other manufacturers may work equally well but have not been tested. Includes: Labeling of BAC clones; Ethanol precipitation; Hybridization; Post-hybridisation treatment / detection. - [Read BAC-FISH Protocol]
Chromatin immunoprecipitation protocol to analyze histone modifications in Arabidopsis thaliana. Werner Aufsatz, Matzke Lab. Gregor Mendel Institute of Molecular Plant Biology, Austria. Includes Chromatin Crosslinking, Chromatin preparation, Pre-clearing and immuno precipitation (IP), Collection,washes and elution of immune complexes, Reverse crosslinking and DNA cleanup - [Read Chromatin immunoprecipitation protocol to analyze histone modifications in Arabidopsis thaliana]
Laboratory sample cleanup is a necessary part for analytical preparation analysis. The removal of Contaminants such as proteins, cell debris and other materials is an important step. Typically this has been done by using Acetonitrile and then Centrifugation to pellet the debris leaving the clean supernant. After this process supernatant can be used for further analysis by HPLC, GC, MS and other analysis tandem methods. HTS Labs. - [Read Protein Precipitation Microplate]
Concentration of DNA by Ethanol Precipitation Protocol. Adapted fromBruce A. Roe, Department of Chemistry and Biochemistry, The University of Oklahoma, Norman, Oklahoma. Usually 2.5 - 3 volumes of ethanol and/or acetate solution is added to the DNA in a microcentrifuge tube. This is then put into an ice-water bath for at least 10 minutes. The precipitation is performed by incubation at -20C overnight. - [Read Concentration of Oligo DNA by Ethanol Precipitation Protocol]
Antigen Design and Sera Purification. Custom antisera. Sigma Aldrich. Peptide Selection and Design, Coupling Strategy
# Selecting the Protein Carrier, Multiple Antigenic Peptides (MAPs), Choice of Host, Adjuvant, Immunization, & Sera Collection, Antisera Purification, Ammonium Sulfate Precipitation, Protein A/G, Immunoaffinity Purification. - [Read Antigen Design and Sera Purification]
This protocol describes the steps required to produce a cDNA microarray. Gene-specific DNA is produced by PCR amplification of purified template plasmid DNAs from cloned ESTs. The PCR product is purified by ethanol precipitation, thoroughly resuspended in - [Read Fabrication Protocol for DNA Microarrays]
General protocol for Ras, Rac, Cdc42, and Rho activation assay. Includes: Affinity Precipitation/Immunoblot Protocol, Cell Culture and Extract Preparation (Adherent and Non Adherent cells), GTPγS/GDP Loading for Positive and Negative Controls, Ras, Rac ,Cdc42, and Rho Pull-Down Assay and Western Blot and Detection. - [Read General Method for Ras, Rac, Cdc42, and Rho Activation Assay]
With this protocol, transcripts that were initiated from specific genes by RNA polymerases prior to permeabilization can be measured. Instead of a nuclear extract, permeabilized cells are used. Includes information on: Permeabilization of Cells; In vitro Transcription Reaction (Run-off); Isolation of RNA; Preparation of Slot Blot Membrane for Hybridization; Hybridization of Nitrocellulose Membrane; TCA Precipitation to Determine Incorporation of [32P] GTP into Nucleic Acid - [Read In Vitro Transcription Assay (Run-off Assay) using Permeabilized Cells]
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.
In Vitro Translated Xenopus Mos Kinase Assay Protocol. In response to progesterone, immature Xenopus oocytes mature to eggs that can be fertilized. The Mos protein kinase is essential for oocyte maturation, most likely due to its ability to activate the MAP kinase cascade. This MAP kinase cascade eventually leads to the activation of Cdc2/cyclin B and entry into M phase. In this protocol, tagged Mos kinase is translated in vitro, immunopurified, and used in a kinase assay.