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Western Blot Home - http://www.molecularstation.com/protein/western-blot/

Category: Protein Protocols: Western Blot Protocol

Western Blot Home. Information on western blotting, western blot procedure and methods, western blot books, stripping protocols. - [Read Western Blot Home]

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Purification of Peroxisomes and Mitochondria from Spinach Leaf by Percoll Gradient Centrifugation - http://www.plantphysiol.org/cgi/reprint/75/3/670

Category: Cell Organelle Protocols: Subcellular Fractionation Protocols: Peroxisome Isolation Protocols

A procedure developed to purify simultaneously peroxisomes and mitochondria from spinach (Spirnacia olercea L.) leaf under isoosmotic and low viscosity conditions. This method involves differential centrifugation and density gradient centrifugation on four layers of Percoll. - [Read Purification of Peroxisomes and Mitochondria from Spinach Leaf by Percoll Gradient Centrifugation]

Quantification made easy - for DNA and Protein Concentration Protocol - http://www.eppendorfna.com/applications/quant_easy.asp?SX=NF&

Category: General Research Protocols and Methods: Spectrophotometry Protocols

Nucleic acid determination. Protein determination Procedure and protocol. Eppendorf. - [Read Quantification made easy - for DNA and Protein Concentration Protocol]

Quantitation of Rare DNAs by PCR Protocol - https://catalog.invitrogen.com/index.cfm?fuseaction=iProtocol.unitSectionTree&treeNodeID=3DD0EEF8E2C50E5631409DE1B105D749&objectid=6676A8D899C49A6D38160C5130F2C612

Category: PCR Protocols: Quantitative PCR Protocols

Protocol that uses the polymerase chain reaction (PCR) to quantitate the numbers of a particular DNA sequence, from 1 to 20,000 molecules per sample. In addition, it helps assess the presence of contaminating sequences, which can seriously affect the outcome of the procedure. - [Read Quantitation of Rare DNAs by PCR Protocol]

Quantitative Telomerase Detection Kit - http://www.biomax.us/pdfs/TelomeraseDetectionKitManual.pdf

Category: Cell Biology and Cell Culture

Quantitative Telomerase Detection Kit. Includes: QTD Real-Time PCR Assay Procedure; Quantitative Telomerase Detection Kit Description. - [Read Quantitative Telomerase Detection Kit]

Radiolabeling of Subtracted cDNA Probes by Random Oligonucleotide Extension Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3860

Category: Nucleic Acid Modification Protocols: Nucleic Acid Radiolabeling Protocols

In this procedure, synthesis of cDNA is carried out in the presence of saturating concentrations of all four dNTPs and trace amounts of a single radiolabeled dNTP. After subtraction hybridization, the enriched single-stranded cDNA is radiolabeled to high specific activity in a second synthetic reaction by extension of random oligonucleotide primers using the Klenow fragment of E. coli DNA polymerase. - [Read Radiolabeling of Subtracted cDNA Probes by Random Oligonucleotide Extension Protocol]

Random Amplification of Polymorphic DNA (RAPD) Typing and Fingerprinting Protocol - http://www.aspergillus.org.uk/indexhome.htm?secure/laboratory_protocols/./rapds.html~main

Category: Fungi Protocols: Aspergillus Protocols

RAPD is a procedure for typing and fingerprinting isolates of a species. It can be used for epidemiological studies, such as investigations into hospital outbreaks and as a laboratory aid to keep track of cultures and to verify that mutants generated in the laboratory are genetically identical to the parental strain. In our hands, the use of one primer, R108, is sufficiently discriminatory to distinguish between the isolates of different strains. - [Read Random Amplification of Polymorphic DNA (RAPD) Typing and Fingerprinting Protocol]

Rat Chromaffin Cells Primary Cultures Standardization and Quality Assessment for Single Cell Assays - http://www.natureprotocols.com/2006/09/29/rat_chromaffin_cells_primary_c.php

Category: Neuroscience Protocols: Neuronal Cell Culture Protocols: Rat Neuronal Cell Culture Protocols

Protocol for standardization of both the isolation procedure and the quality of the cultures. - [Read Rat Chromaffin Cells Primary Cultures Standardization and Quality Assessment for Single Cell Assays]

Rat Chromaffin Cells Primary Cultures: Standardization and Quality Assessment for Single-Cell Assay - http://www.natureprotocols.com/2006/09/29/rat_chromaffin_cells_primary_c.php

Category: Primary Cell Isolation and Culture Protocols: Mammalian Cell Culture Protocols

Protocol to be used as reference to help standardization of both the isolation procedure and the quality of the cultures allowing a better comparison of the results obtained from different laboratories. - [Read Rat Chromaffin Cells Primary Cultures: Standardization and Quality Assessment for Single-Cell Assay]

Reverse Ligation Mediated RT - PCR - http://www.liv.ac.uk/~giles/methods/rlpcr.htm

Category: RNA Protocols: RT-PCR and cDNA synthesis

This procedure was first described by Bertrand et al to demonstrate that ribozymes could be enzymatically active in vivo. We adapted the method to show that certain oligodeoxynucleotides could direct the activity of endogenous ribonuclease H to cleave tar - [Read Reverse Ligation Mediated RT - PCR]

RNA Labeling by In Vitro Transcription of DNA with DIG, Biotin or Fluorescein RNA Labeling Mix - http://www.roche-applied-science.com/PROD_INF/MANUALS/InSitu/pdf/ISH_54-57.pdf

Category: Cytogenetics Protocols: RNA In Situ Hybridization Protocols

Protocol for RNA labeling by in vitro transcription of DNA with DIG, Biotin or Fluorescein RNA Labeling Mix. A PCR fragment that has the appropriate promoter ligated to its 5’-ends can also serve as a transcription template. The procedure described incorporates one modified nucleotide (DIG-, Biotin-, or Fluorescein-UTP) at approximately every 20 – 25th position in the transcripts. - [Read RNA Labeling by In Vitro Transcription of DNA with DIG, Biotin or Fluorescein RNA Labeling Mix]

RNAi Screens in C. elegans Protocol - http://www.natureprotocols.com/2006/07/19/rnai_screens_in_c_elegans_in_a.php

Category: RNAi Technology Protocols: RNAi C. elegans Protocols

Protocol for RNAi screens in C. elegans in a 96-well liquid format and their application to the systematic identification of genetic interactions. The procedure allows thousands of RNAi feeding experiments to be performed per investigator per day. - [Read RNAi Screens in C. elegans Protocol]

Sampling of Aspergillus spp Spores in Air Protocol - http://www.aspergillus.org.uk/indexhome.htm?secure/laboratory_protocols/./airprotocol.html~main

Category: Fungi Protocols: Aspergillus Protocols

Protocol allows the isolation and enumerate Aspergillus spp spores in air. Includes: Sampling Procedure; Sampling Location; Selection of Sampling Time; Sampling Steps; Laboratory Procedure; Enumerating the Colony Forming Units. - [Read Sampling of Aspergillus spp Spores in Air Protocol]

Sandwich ELISA Procedure - http://research.cm.utexas.edu/bkitto/Kittolabpage/Protocols/Biochemistry/sELISA.html

Category: ELISA Protocols: Sandwich ELISA Assay Protocols

Sandwich ELISA Procedure. Kitto Lab, Univ. Texas at Austin - [Read Sandwich ELISA Procedure]

Screening Bacterial Colonies by Hybridization: Large Numbers Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3937

Category: Bacterial Protocols

This procedure is used to plate, replicate, and subsequently screen large numbers of bacterial colonies (up to 2 x 104 colonies per 150-mm plate or 104 colonies per 90-mm plate). - [Read Screening Bacterial Colonies by Hybridization: Large Numbers Protocol]

Silver Staining of Gels for Mass Spectrometry - http://www.healthsystem.virginia.edu/internet/biomolec/Silverstains.cfm

Category: Mass Spectrometry Protocols

Silver Staining of Gels for Mass Spectrometry. Desiderio procedure: R.R. Becklin. Univ. Virginia. - [Read Silver Staining of Gels for Mass Spectrometry]

Simple Western Blot Protocol - http://www.prosci-inc.com/Antibody/Western-Blot-Procedure.html

Category: Protein Protocols: Western Blot Protocol

Simple Western Blotting Protocol ProSci. - [Read Simple Western Blot Protocol]

Simple, Reliable Steps for DNA Fragment Isolation and Purification Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/16/pdb.prot4389

Category: Mouse Protocols: Mouse Genetics Protocols

Protocol describes a simple procedure for DNA fragment isolation and purification. The resulting DNA fragment can then. be used for microinjection to produce transgenic mice - [Read Simple, Reliable Steps for DNA Fragment Isolation and Purification Protocol]

Single AP staining Protocol - http://icg.cpmc.columbia.edu/cattoretti/Protocol/immunohistochemistry/nbtbcipstaining.html#PROCEDURE

Category: Histology Protocols: Histology Staining Procedures

Protocol for single AP staining. - [Read Single AP staining Protocol]

Single-tube Coupled Transcription/Translation Reactions for Eukaryotic In Vitro - http://www.promega.com/tbs/tm045/tm045.pdf

Category: Cell Biology and Cell Culture: In Vitro Reconstitution Protocols

Information for protocol using single-tube, coupled transcription/translation reactions for eukaryotic in vitro translation. Includes information on: Translation Procedure; Positive Control Translation Reactions Using Luciferase; Cotranslational Processing Using Canine Pancreatic Microsomal Membranes; Post-Translational Analysis; Positive Control Luciferase Assays; Composition of Buffers and Solutions; Luciferase SP6/T7 Control DNAs - [Read Single-tube Coupled Transcription/Translation Reactions for Eukaryotic In Vitro]

Spore Germination Protocol - http://www-rcf.usc.edu/~forsburg/spores.html

Category: Yeast Cell Culture Protocols: Yeast Sporulation Protocols

Protocol for spore germination. This procedure is typically used for the isolation and preparation of spores from a diploid strain heterozygous for a marked disruption (e.g., yfg1::his3+) Inoculation of the spore population into minimal medium lacking the nutritional supplement corresponding to the disruption marker (e.g., minimal medium lacking histidine) allows only the disruption spores to germinate. - [Read Spore Germination Protocol]

Stable Isotope Labeling by Amino Acids in Cell Culture SILAC Protocol - http://www.natureprotocols.com/2007/01/11/a_practical_recipe_for_stable.php

Category: Chromatography Protocols: Liquid Chromatography Protocols

Protocol for stable isotope labeling by amino acids in cell culture (SILAC). Protocol describes how to apply SILAC and the use of nano-scale liquid chromatography coupled to electrospray ionization mass spectrometry for protein identification and quantification. This procedure can be completed in 8 days. - [Read Stable Isotope Labeling by Amino Acids in Cell Culture SILAC Protocol]

TCA/Acetone Precipitation (Large Scale) PDF - http://www.signaling-gateway.org/data/cgi-bin/ProtocolFile.cgi/afcs_PP00000148.pdf?pid=PP00000148

Category: Protein Protocols: Protein Precipitation Procols

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]

Tetracycline as Regulator of Inducible Gene Expression III - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3958

Category: Transfection Protocols: Stable Transfection Protocols

Stably transfected cells, generated in the first two stages of the procedure, are induced for expression of the target gene. After harvesting and lysis, the lysates are analyzed by SDS-PAGE and immunoblotting. - [Read Tetracycline as Regulator of Inducible Gene Expression III]

Tetracycline as Regulator of Inducible Gene Expression Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3986

Category: Transfection Protocols: Stable Transfection Protocols

Protocol uses an autoregulatory system in which the transcriptional trans-activator tTA drives its own expression and that of a target gene. The first stage of the procedure describes how to generate stable lines of NIH-3T3 cells that express either tTA alone or tTA and the tetracycline-regulated target gene. - [Read Tetracycline as Regulator of Inducible Gene Expression Protocol]

Tetracycline as Regulator of Inducible Gene Expression Protocol II - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3957

Category: Transfection Protocols: Stable Transfection Protocols

This stage of the procedure describes the transfection with target genes of cell lines already expressing inducible tTA. In this example, the target genes are transfected on a plasmid that carries puromycin resistance as a selectable marker. - [Read Tetracycline as Regulator of Inducible Gene Expression Protocol II]

Articles (1-10 of 12)

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.