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Isolation of poly A+ RNA from Total RNA by Oligo(deoxythymidine)cellulose Chromatography - http://wheat.pw.usda.gov/~lazo/methods/au/m6e1.html

Category: Chromatography Protocols: DNA RNA Affinity Chromatography

Isolation of poly A+ RNA from Total RNA by oligo(deoxythymidine)cellulose Chromatography Protocol. Faulkner-Jones - [Read Isolation of poly A+ RNA from Total RNA by Oligo(deoxythymidine)cellulose Chromatography]

Isolation of Total RNA form E. coli for Microarrays Protocol - http://bugarrays.stanford.edu/protocols/rna/Total_RNA_from_Ecoli.pdf

Category: E Coli Protocols: E coli Nucleic Acid Protocols

Protocol for the isolation of total RNA from E. coli for Microarrays. Includes: Harvesting total RNA; DNase I treatment of RNA samples. - [Read Isolation of Total RNA form E. coli for Microarrays Protocol]

Isolation of Total RNA From E. Coli for Microarrays Protocol - http://bugarrays.stanford.edu/protocols/rna/Total_RNA_from_Ecoli.pdf

Category: Microarray Protocols: Microarray Analysis Protocols

Protocol for isolation of total RNA from E. coli for microarrays. Includes: Harvesting total RNA; Dnase I treatment of RNA samples. - [Read Isolation of Total RNA From E. Coli for Microarrays Protocol]

Laser-capture Microdissection Protocol - http://www.nature.com/nprot/journal/v1/n2/full/nprot.2006.85.html#a1

Category: Primary Cell Isolation and Culture Protocols: Laser Capture Microdissection Protocols

LCM technology can harvest the cells of interest directly or can isolate specific cells by cutting away unwanted cells to give histologically pure enriched cell populations. A variety of downstream applications exist: DNA genotyping and loss-of-heterozygosity (LOH) analysis, etc. Protocol provides a thorough description of LCM techniques, with an emphasis on tips and troubleshooting advice derived from LCM users. The total time required to carry out this protocol is typically 1–1.5 h. - [Read Laser-capture Microdissection Protocol]

Lowry â€“ Protein Determination - http://wolfson.huji.ac.il/purification/Protocols/loury.html

Category: Protein Protocols: Lowry Protein Assay

Lowry â€“ Protein Determination. Quantitation of total protein using the lowry protein assay. Dr. Mario Lebendiker. The Wolfson Centre for Applied Structural Biology, The Hebrew University of Jerusalem. Method materials and notes for the Lowry assay. - [Read Lowry â€“ Protein Determination]

Lung Cell Assay Protocol - http://embryo.ib.amwaw.edu.pl/invittox/prot/48.htm

Category: Pharmacology and Toxicology Protocols: Cytotoxicity Protocols: Cell Specific Cytotoxicity Assays Protocols

Potential embryotoxicity is assessed by monitoring the effect of the test compound on total protein synthesis, and DNA synthesis in cultured human foetal lung fibroblasts. Rat lung epithelial cells can be used to determine cytotoxicity of select compounds because of their ability to metabolise xenobiotics. - [Read Lung Cell Assay Protocol]

Lysis and Protein Extraction from WEHI-231 Cells with TriPure Isolation Reagent Protocol - http://www.signaling-gateway.org/data/cgi-bin/ProtocolFile.cgi/afcs_PP00000162.pdf?pid=PP00000162

Category: Chromatography Protocols: Immobilized Metal Ion Affinity Chromatography Protocols

Protocol is the first step in a three-step process for the preparation and enrichment of phosphopeptides using immobilized metal affinity chromatography (IMAC) for the identification of the phosphopeptides by LC-MS/MS. This procedure is used to prepare protein extracts from WEHI-231 cells. This preparation method provides total cellular protein samples that are free of contaminating nucleic acids. - [Read Lysis and Protein Extraction from WEHI-231 Cells with TriPure Isolation Reagent Protocol]

Methods for Plant RNA Isolation - http://wheat.pw.usda.gov/~lazo/methods/lazo/met6.html

Category: Plant Biology Protocols: Plant DNA RNA Isolation Protocols

AMES/Chloroform extraction to extract total RNA from potato leaves for viroid analysis. - [Read Methods for Plant RNA Isolation]

Microarray Protocol for Affymetrix In Situ Synthesized Oligo Arrays - http://www.signaling-gateway.org/data/cgi-bin/ProtocolFile.cgi/afcs_PP00000174.pdf?pid=PP00000174

Category: Microarray Protocols: Microarray Hybridization Protocols

Protocol details the preparation of biotin-labeled target samples and hybridization of these samples to an Affymetrix in situ synthesized oligonucleotide GeneChip array. The procedure requires a minimum of 5 µg of purified total RNA as starting material. - [Read Microarray Protocol for Affymetrix In Situ Synthesized Oligo Arrays]

Microarray Protocol for Agilent Inkjet-Deposited Presynthesized - http://www.signaling-gateway.org/data/cgi-bin/ProtocolFile.cgi/afcs_PP00000184.pdf?pid=PP00000184

Category: Microarray Protocols: Amino-allyl Microarray Methods

Protocol details the preparation of fluorescently labeled target samples (aminoallyl method) and hybridization of these samples to a microarray of Agilent inkjet-deposited presynthesized oligonucleotides. The procedure requires a minimum of 3 µg of purified total RNA as starting material. - [Read Microarray Protocol for Agilent Inkjet-Deposited Presynthesized]

Microarray Protocol for Agilent Inkjet-Deposited Presynthesized Oligo Array - http://www.signaling-gateway.org/data/cgi-bin/ProtocolFile.cgi/afcs_PP00000179.pdf?pid=PP00000179

Category: Microarray Protocols: Microarray DNA Labeling Protocols

Protocol details the preparation of fluorescently labeled target samples and hybridization of these samples to a microarray of Agilent inkjet-deposited presynthesized oligonucleotides. The procedure requires a minimum of 3 µg of purified total RNA as starting material. - [Read Microarray Protocol for Agilent Inkjet-Deposited Presynthesized Oligo Array]

Microarray Protocol for Agilent Inkjet-Deposited Presynthesized Oligo Arrays - http://www.signaling-gateway.org/data/cgi-bin/ProtocolFile.cgi/afcs_PP00000179.pdf?pid=PP00000179

Category: Microarray Protocols: Microarray Hybridization Protocols

Protocol details the preparation of fluorescently labeled target samples and hybridization of these samples to a microarray of Agilent inkjet-deposited presynthesized oligonucleotides. The procedure requires a minimum of 3 µg of purified total RNA as starting material. Includes: cDNA Synthesis; Fluorescent cRNA Synthesis; cRNA Precipitation and Cleanup; cRNA Quantification; Hybridization; Washing. - [Read Microarray Protocol for Agilent Inkjet-Deposited Presynthesized Oligo Arrays]

Microinjection of Mouse ES Cells into Blastocysts - http://www.bcm.edu/dtmc/?pmid=7791

Category: Microinjection Protocols: Microinjection of DNA Protocols

The core defines one injection as a total of forty blastocysts injected on two consecutive days using one or two clones for the same mutation. Please, refer to the Pricing page for information on the cost of one injection. - [Read Microinjection of Mouse ES Cells into Blastocysts]

Neutral Red Cytotoxicity Assay Protocol - http://embryo.ib.amwaw.edu.pl/invittox/prot/64.htm

Category: Pharmacology and Toxicology Protocols: Cytotoxicity Protocols: Cytotoxicity Assays in Cell Culture Protocols

The cytotoxic effect of chemicals upon mammalian cells, such as BALB/c 3T3 and HepG2, in culture is measured by highest tolerated dose (HTD), cell viability (Neutral Red) and total cell protein (coomassie blue). - [Read Neutral Red Cytotoxicity Assay Protocol]

Preparation of B-Lymphocyte RNA for Microarray Analysis Protocol - http://www.signaling-gateway.org/data/cgi-bin/ProtocolFile.cgi/afcs_PP00000009.pdf?pid=PP00000009

Category: Immunology: B Cell Protocols

Procedure permits the isolation of at least 5 µg of total RNA from a sample of purified mouse splenic B lymphocytes. The quality of the RNA is assessed by separation of an aliquot through 1% agarose and staining with ethidium bromide as described in AfCS protocol Visualization of RNA Preparations on 1% Agarose Gels. The isolated RNA is used for analysis of gene expression by microarray technology. analysis of gene expression by microarray technology. - [Read Preparation of B-Lymphocyte RNA for Microarray Analysis Protocol]

Preparation of B-Lymphocyte RNA for Microarray Analysis Protocol - http://www.signaling-gateway.org/data/cgi-bin/ProtocolFile.cgi/afcs_PP00000009.pdf?pid=PP00000009

Category: Microarray Protocols: Microarray Analysis Protocols

Protocol permits the isolation of at least 5 µg of total RNA from a sample of purified mouse splenic B lymphocytes Isolation of Resting B Lymphocytes from Sixteen Mouse Spleens. - [Read Preparation of B-Lymphocyte RNA for Microarray Analysis Protocol]

Preparation of Fluorescent DNA Probe from HUMAN mRNA or Total RNA using Direct Incorporation - http://cmgm.stanford.edu/pbrown/protocols/5_hyb_human.html

Category: Nucleic Acid Detection Protocols

Protocol for preparation of fluorescent DNA probe from HUMAN mRNA or total RNA using direct incorporation. Includes: Preparing fluoresenctly labeled cDNA (probe); Washing and Scanning Arrays. - [Read Preparation of Fluorescent DNA Probe from HUMAN mRNA or Total RNA using Direct Incorporation]

Preparation of Fluorescent DNA Probe from HUMAN mRNA/ Total RNA using Direct Incorporation Protocol - http://cmgm.stanford.edu/pbrown/protocols/5_hyb_human.html

Category: Microarray Protocols: Microarray Hybridization Protocols

Protocol for the preparation of fluorescent DNA probe from HUMAN mRNA or total RNA using direct incorporation. Includes: Preparing fluoresenctly labeled cDNA (probe); Washing and Scanning Arrays. - [Read Preparation of Fluorescent DNA Probe from HUMAN mRNA/ Total RNA using Direct Incorporation Protocol]

Preparation of Myocyte RNA for Microarray Analysis Protocol - http://www.signaling-gateway.org/data/cgi-bin/ProtocolFile.cgi/afcs_PP00000133.pdf?pid=PP00000133

Category: Microarray Protocols: Microarray Analysis Protocols

Protocol permits the isolation of at least 3 µg of total RNA from approximately 150,000 cultured cardiac myocytes from adult mice. Includes: Treatment of Samples and Termination of Reactions; Isolation of RNA; Use of Environmental Chamber. - [Read Preparation of Myocyte RNA for Microarray Analysis Protocol]

Processing of Microdissected Tissue for Molecular Analysis RNA-based Studies - http://cgap-mf.nih.gov/Protocols/Processing/RNA.html

Category: Primary Cell Isolation and Culture Protocols: Laser Capture Microdissection Protocols

RNA-based studies of processing of microdissected tissue for molecular analysis. Includes: Total RNA Isolation; Optional DNAse Treatment. - [Read Processing of Microdissected Tissue for Molecular Analysis RNA-based Studies]

Protein Trafficking Methods - http://mips.gsf.de/proj/eurofan/eurofan_2/n5/#Methods

Category: Protein Protocols: Protein Trafficking

Methods on protein trafficking. Information on N5 Secretion and protein trafficking. Methods include: Alcian Blue Test; Lucifer Yellow uptake assay; Preparation of total protein extracts for western immunoblots; Screen for Drug Sensitivity. - [Read Protein Trafficking Methods]

Protocols for microarray development in use in the Vodkin Laboratory - http://soybeangenomics.cropsci.uiuc.edu/protocols/index.html

Category: DNA Microarray Protocols

Reverse transcriptase labeling with Cy3/Cy5, Staining of total DNA on slides, construction, Laser scanning, Spot quantitation, Statistics, sample tracking. Vodkin Laboratory. U.Illinois. - [Read Protocols for microarray development in use in the Vodkin Laboratory]

Purification of poly(A)+ RNA from total RNA - http://kuchem.kyoto-u.ac.jp/seika/shiraishi/protocols/oligotex.html

Category: RNA Protocols: mRNA Extraction Protocols

Purification of poly(A)+ RNA from total RNA. Hideaki Shiraishi, Kyoto University. The protocol employs Oligotex-dT30 for poly(A)+ RNA purification. - [Read Purification of poly(A)+ RNA from total RNA]

Purification of RNA from Cells and Tissues by Acid Phenol-Guanidinium Thiocyanate-Chloroform Extract - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot4045

Category: Nucleic Acid Purification Protocols: RNA Isolation Protocols: RNA Isolation fromTissues or Cultured Cells Protocols

Single-step technique, cells are homogenized in guanidnium thiocyanate and the RNA is purified from the lysate by extraction with phenol:chloroform at reduced pH. Many samples can be processed simultaneously and speedily. The yield of total RNA depends on the tissue or cell source and is generally in the range of 4-7 µg/ml starting tissue or 5-10 µg/106 cells. IMPORTANT: Prepare all reagents used in this protocol with Diethyl pyrocarbonate (DEPC)-treated H2O. - [Read Purification of RNA from Cells and Tissues by Acid Phenol-Guanidinium Thiocyanate-Chloroform Extract]

Rapid and Reliable Preparation of Total Cell Lysates from Fission Yeast Protocol - http://www.natureprotocols.com/2006/07/21/rapid_and_reliable_preparation.php

Category: Yeast Protocols: Yeast Protein Protocols: Yeast Protein Purification and Isolation Protocols

Protocol for rapid and reliable preparation of total cell lysates from fission yeast. Method for successful and reproducible extraction of proteins from S. pombe cells. - [Read Rapid and Reliable Preparation of Total Cell Lysates from Fission Yeast Protocol]

Articles (1-10 of 11)

Populational Analysis of Genomic DNA Protocol

A protocol on the populational Analysis of genomic DNA.

Lambda Phage Protocol for Large Preparation DNA Recovery

Lambda Phage Protocol for Large Preparations DNA Recovery.

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.

Microarray Protocol Preparation of Fluorescent DNA Probes from Human mRNA

This Microarray Protocol Preparation of Fluorescent DNA Probes from Human mRNA protocol describes the production of probes labeled with the fluorescent dyes, Cy3 and Cy5, following the synthesis of cDNA from human mRNA and the hybridization of the probes to DNA microarrays.

In Vitro Translated Xenopus Mos Kinase Assay Protocol

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.

Immobilized Antigen Phage Antibody Selection Protocol

A protocol for the selection of Phage Antibodies using Immobilized Antigen. This method describes the selection of antibodies from bacteriophage antibody libraries that recognize a specific antigen. The phage display library of antibody-displaying phage particles is exposed to antigen attached to a solid substrate (Nunc Immuno™ tubes). The phage particles with affinity for antigen bind to the immobilized antigen and are selected from the library of phage expressing antibodies.

Constructing Random-Peptide Libraries for Phage Display

The protocol describes the construction of a large (10^8 to 10^10) primary random peptide phage library in fUSE5 or f88-4 strains.

Coupling Cysteine Containing Peptides to Carrier Protein for Immunization and Polyclonal Antibody

This protocol is used in the production of polyclonal antibodies that recognize a peptide sequence of interest.

3' Rapid Amplification of cDNA Ends RACE Using PCR Protocol

3' Rapid Amplification of cDNA Ends RACE Using PCR Protocol. This protocol contains the steps for 3' end rapid amplification of mRNA by PCR. The first-strand cDNA is synthesized from total or poly(A+) RNA by priming from the poly-A tail of the mRNA using a oligo (dT) adaptor primer. The cDNA is then amplified via PCR using a gene-specific primer and an adaptor primer.

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