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70% Ethanol Fixation for Recovery of DNA, RNA, and Protein Protocol - http://cgap-mf.nih.gov/Protocols/Tissues/TissueProtocols/EthanolFixation.html

Category: Primary Cell Isolation and Culture Protocols: Laser Capture Microdissection Protocols: Tissue Preparation for LCM Protocols

Protocol for 70% ethanol fixation for recovery of DNA, RNA, and protein. - [Read 70% Ethanol Fixation for Recovery of DNA, RNA, and Protein Protocol]

70% Ethanol Fixation for Recovery of DNA, RNA, and Protein Protocol - http://cgap-mf.nih.gov/Protocols/Tissues/TissueProtocols/EthanolFixation.html

Category: Primary Cell Isolation and Culture Protocols

Protocol for 70% ethanol fixation for recovery of DNA, RNA and protein. - [Read 70% Ethanol Fixation for Recovery of DNA, RNA, and Protein Protocol]

Analysis of DNA Content and Surface Immunophenotype Protocol Using Ethanol Fixation - http://www.cyto.purdue.edu/flowcyt/research/cytotech/telford/ethanol.htm

Category: Immunohistochemistry Protocols: Fixation Protocols

Protocol for analysis of DNA content and surface immunophenotype using gentle ethanol fixation techniques. - [Read Analysis of DNA Content and Surface Immunophenotype Protocol Using Ethanol Fixation]

Analysis of DNA Fragmentation Using Propidium Iodide (PI) Staining After Ethanol Fixation Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4431

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

This protocol provides a method for analysis of DNA fragmentation using flow cytometry after propidium iodide (PI) labeling of fixed apoptotic cells. Flow cytometry reveals apoptotic nuclei in the subdiploid region of the cell cycle histogram... - [Read Analysis of DNA Fragmentation Using Propidium Iodide (PI) Staining After Ethanol Fixation Protocol]

BAC-FISH Protocol - http://www.nhm.ac.uk/hosted_sites/schisto/protocols/BAC_FISH.html

Category: BACs Bacterial Artificial Chromosome Protocols

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]

Clearing Arabidopsis Non-Ovule Materials With the HCG Solution Protocol - http://www.hybtech.org/Liu/HCG_clearing.htm

Category: Plant Biology Protocols

Protocol can be used for clearing intact non-ovule materials of arabidopsis, which can then be observed under Normarski optics. This is an efficient way to analyse root, seedling even flower development without sectioning. This protocol could also be used for clearing GUS stained material, after chlorophyll is removed by 70% ethanol. - [Read Clearing Arabidopsis Non-Ovule Materials With the HCG Solution Protocol]

Concentrating and Desalting Nucleic Acids with Microconcentrators Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4457

Category: Nucleic Acid Purification Protocols

Ultrafiltration is an alternative to ethanol precipitation for the concentration and desalting of nucleic acid solutions. It requires no phase change and is particularly useful for dealing with very low concentrations of nucleic acids. This protocol describes the use of the Microcon cartridge, a centrifugal ultrafiltration device, to concentrate and desalt nucleic acid solutions. - [Read Concentrating and Desalting Nucleic Acids with Microconcentrators Protocol]

Concentration of Oligo DNA by Ethanol Precipitation Protocol - http://mycoplasmas.vm.iastate.edu/lab_site/methods/DNA/precip.html

Category: Oligonucleotide Protocols: Oligonucleotide Purification Protocols

Concentration of DNA by Ethanol Precipitation Protocol. Adapted from Bruce 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]

DNA cloning Procedures - http://hulab.tamu.edu/Protocols/cloning/

Category: DNA Protocols: DNA Cloning

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]

Fabrication Protocol for DNA Microarrays - http://research.nhgri.nih.gov/microarray/fabrication.shtml

Category: DNA Microarray Protocols

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]

FIXATION and DNA Staining for Cell Cycle Analysis - http://www.biotech.iastate.edu/facilities/CELLHYB/CCAnalysis.htm

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols

FIXATION and DNA Staining for Cell Cycle Analysis Protocol. This method of DNA staining utilizes ethanol to fix the cells and permeabilize the membrane, which allows the dye (Propidium Iodide) to enter the cells. Propidium Iodide (PI) is a DNA-binding fluorochrome that intercalates in the double-helix. Ribonuclease-A is used to eliminate the staining of double-stranded RNA. - [Read FIXATION and DNA Staining for Cell Cycle Analysis]

Measurement of DNA Content Using Propidium Iodide (PI) Staining of Fixed Whole Cells Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4436

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

This protocol describes a method for quantitative measurement of DNA using propidium iodide (PI) staining and flow cytometry. PI stains all double-stranded regions of both DNA and RNA by intercalating between the stacked bases of the double helix. PI cannot penetrate an intact cell membrane; therefore, cells are fixed prior to staining. The ethanol-fixed cells can be stored unstained at 4°C for days, or even weeks, and then stained and analyzed. - [Read Measurement of DNA Content Using Propidium Iodide (PI) Staining of Fixed Whole Cells Protocol]

Measurement of GFP Expression and DNA Content in Permeabilized Cells - http://cyto.mednet.ucla.edu/Protocols/DNA%20and%20GFP%20PI.htm

Category: Reporter Gene Assays: GFP Assay Protocols

Protocol measuring the amount of GFP expression and DNA content in permeabilized cells. Includes: Fix cells with formaldehyde; Permeabilize cells with ethanol; Preparation of Buffered 2% Formaldehyde Solution. - [Read Measurement of GFP Expression and DNA Content in Permeabilized Cells]

Measurement of GFP Expression and DNA Content in Permeabilized Cells Protocol - http://cyto.mednet.ucla.edu/Protocols/DNA%20and%20GFP%20PI.htm

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

Protocol for the measurement of GFP expression and DNA content in permeabilized cells. Includes: Fix cells with formaldehyde; Permeabilize cells with ethanol; Preparation of Buffered 2% Formaldehyde Solution. - [Read Measurement of GFP Expression and DNA Content in Permeabilized Cells Protocol]

Preparation of Single-stranded Bacteriophage M13 DNA Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3686

Category: Viral Manipulation Protocols: M13 Phage Manipulation Protocols

Bacteriophage M13 single-stranded DNA is prepared from virus particles secreted by infected cells into the surrounding medium. The filamentous particles are concentrated by precipitation from a high-ionic-strength buffer with polyethylene glycol. Subsequent extraction with phenol releases the single-stranded DNA, which is then collected by precipitation with ethanol. This protocol is generally used to prepare single-stranded DNA from a small number of M13 isolates. - [Read Preparation of Single-stranded Bacteriophage M13 DNA Protocol]

Protein Precipitation Protocols - http://www.ls.huji.ac.il/~purification/Protocols/ProteinPrecipitation.html

Category: Protein Protocols: Protein Precipitation Procols

TCA-DOC Normal, TCA Acetone Precipitation, Ethanol Precipitation, TCA-DOC / Acetone, Acidified Acetone/Methanol, Dr. Mario Lebendiker. University of Jerusalem. - [Read Protein Precipitation Protocols]

Recovery of DNA from Low-melting-temperature Agarose Gels: Enzymatic Digestion with Agarase Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot4026

Category: Nucleic Acid Electrophoresis Protocols: DNA Electrophoresis Protocols: DNA Electrophoresis in Agarose Gels Protocols

A fragment of gel containing a band of DNA is excised and digested with agarase, which hydrolyzes the polymer to disaccharide subunits. The released DNA is then purified by phenol extraction and ethanol precipitation. The method works well for DNAs ranging in size from <5 kb to >20 kb. - [Read Recovery of DNA from Low-melting-temperature Agarose Gels: Enzymatic Digestion with Agarase Protocol]

Simultaneous Analysis of DNA Content and Surface Immunophenotype Protocol - http://www.cyto.purdue.edu/flowcyt/research/cytotech/telford/ethanol.htm

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]

Standard Ethanol Precipitation of DNA in Microcentrifuge Tubes Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4456

Category: Nucleic Acid Purification Protocols

Protocol describes the standard method to recover nucleic acids from aqueous solutions by precipitation of DNA with ethanol. Subnanogram amounts of DNA (and RNA) can be quantitatively precipitated with ethanol, collected by centrifugation, and redissolved within minutes. - [Read Standard Ethanol Precipitation of DNA in Microcentrifuge Tubes Protocol]

Articles (1-9 of 9)

Populational Analysis of Genomic DNA Protocol

A protocol on the populational Analysis of genomic DNA.

Single Stranded Plasmid DNA Isolation Protocol

A Single Stranded Plasmid DNA Isolation Protocol describing the production and isolation of single-stranded DNA (ssDNA) using bacteriophagemid-containing bacteria and helper phage. Infection of the host cells with helper phage allows for packaging of ssDNA into bacteriophage. The ssDNA can then be isolated from phage particles.

Genomic DNA Labeling of Microarrays 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.

Acid Guanidinium Thiocyanate RNA Isolation Phenol Chloroform Extraction

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.

Paraffin Embedding Protocol

Paraffin Embedding Protocol for molecular profiling. This Paraffin Embedding Protocol describes the processing of the tissues into sections following ethanol fixation. Molecular profiling (MP) is a technique that is used to visualize the global patterns of RNA expression or protein expression in various cell types and disease processes.

DNA Fragment Purification from Polyacrylamide Gels Protocol

A simple protocol for the purification of DNA fragments from Polyacrylamide Gels.

Absorption Spectroscopy and Quantification of Filamentous Phage Protocol

Unlike spherical phage, such as T4 and λ, which have roughly equal weight ratios of protein to DNA, filamentous phage have about six times more protein than DNA; the protein therefore contributes substantially to the absorption spectrum.

Picking Transfected ES Cell Clones Protocol

This protocol a protocol on how to generate transfected embryonic stem (ES) cell clones. The previous protocol in this series is the Protocol for Electroporation of ES cells. The next protocol in the series is the Protocol on Disaggregation, Expansion, and Freezing of Transfected ES Clones.

Electrotransformation of BMH 81-17mut S for Isolating Site-Directed dsDNA Mutants

This protocol describes the electroporation of the BMH 81-17 mut S strain that is recommended for tranformation of the site directed mutagenesis of dsDNA (See Protocol on Site-Directed Mutagenesis on Double Stranded DNA). BMH 81-17 mut S are a mismatch repair defective (mut S) Escherichia coli strain. The probability that the two mutations will cosegregate during the first round of DNA replication is increased in this strain.

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