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MONOCYTE AND NEUTROPHIL ISOLATION - http://aactg.s-3.com/pub/download/imm/phagocyt.pdf

MONOCYTE AND NEUTROPHIL ISOLATION. The protocol describes the method used for isolation of monocytes and neutrophils used in both the chemotaxis assay and microbicidal assays. Pediatric AIDS Clinical Trials Group - [Read MONOCYTE AND NEUTROPHIL ISOLATION]

mRNA Preparation - http://www.promega.com/tbs/tm021/tm021.html#iv

Category: RNA Protocols: mRNA Extraction Protocols

mRNA Preparation. Promega. Abstract for PolyATtractÂ® mRNA Isolation Systems - [Read mRNA Preparation]

Neurospora Methods Protocols - http://www.fgsc.net/methods/stanford.html

Category: Fungi Protocols: Neurospora Protocols

Protocols for Neurospora methods. Includes: Standard strains; Crosses; Minimal medium; Color coding of media; Agar substrate for manipulation and isolation; Stock solutions of supplements; Mating-type tests; Preservation of stocks by silica gel; Cleaning of glassware; Control of mites. - [Read Neurospora Methods Protocols]

Northern Blot Analysis of mRNA From Mammalian Polyribosomes Protocol - http://www.natureprotocols.com/2006/09/15/northern_blot_analysis_of_mrna.php

Category: Nucleic Acid Detection Protocols: Nucleic Acid Blotting Protocols

The protocol consists of a method for the generation of cytoplasmic extracts from mammalian cells (in this case, 293T cells) without the disruption of polyribosomes, the separation of ribosomal components and polyribosomes by sucrose gradient centrifugation, the isolation of mRNA from these fractions, and detection of mRNA by Northern blot analysis. - [Read Northern Blot Analysis of mRNA From Mammalian Polyribosomes Protocol]

Nuclei Isolation Protocol - http://wheat.pw.usda.gov/~lazo/methods/lazo/dnaplnuc.html

Category: Plant Biology Protocols: Plant DNA RNA Isolation Protocols

Protocol for nuclei isolation. - [Read Nuclei Isolation Protocol]

Nucleolar Isolation Protocol - http://www.lamondlab.com/f7nucleolarprotocol.htm

Category: Cell Organelle Protocols: Nucleus Protocols

Protocol for nuclear isolation. - [Read Nucleolar Isolation Protocol]

One-Step Isolation of Plant DNA Suitable for PCR Amplification - http://pubs.nrc-cnrc.gc.ca/ispmb/ispmb19/r01-054.pdf

Category: Plant Biology Protocols: Plant DNA RNA Isolation Protocols

One step extraction for isolation of plant DNA. DNA suitable for amplification by PCR can be produced from leaf material smaller than 0.3 mm2 in less than 20 min & no tube changes. Method was tested on several plant species. Method was found to extract DNA that could be amplified without any further purification or treatment. The isolated DNA was amplified using a universal chloroplast primer set. The method was validated by comparing size of PCR products generated using standard DNA isolation. - [Read One-Step Isolation of Plant DNA Suitable for PCR Amplification]

Phenol-based Protocol Isolation of DNA Fragments from Low-Melting Temperature Agarose - http://wheat.pw.usda.gov/~lazo/methods/101/101lmtagar.html

Category: DNA Protocols: DNA Extraction Protocols: Agarose Gel DNA Extraction Protocols

Phenol-based Protocol Isolation of DNA Fragments from Low-Melting Temperature Agarose. Reference: Favre, D. 1992. Biotechniques vol. 13. - [Read Phenol-based Protocol Isolation of DNA Fragments from Low-Melting Temperature Agarose]

Picking Single Phage Plaques Protocol - http://humgen.wustl.edu/hdk_lab_manual/lambda/lambda1.html

Category: Viral Culture and Isolation Protocols: Viral Culture Protocols

Protocol for the isolation of single bacteriophage lambda plaques to be used for making lysates. - [Read Picking Single Phage Plaques Protocol]

Plant Protein Isolation - http://www.oardc.ohio-state.edu/stockingerlab/Protocols/ProteinIsolation.pdf

Category: Protein Protocols: Protein Extraction From Tissue

Plant Protein Isolation - Rapid isolation of protein for SDS-PAGE analysis. (Essentially the same protocol as that described for GUS Assays). Stockinger Lab - [Read Plant Protein Isolation]

Plasmid Isolation and Analysis. Pictorial Protocol. - http://www.biotech.iastate.edu/publications/ppt_presentations/html/IsoPart1_files/v3_document.htm

Category: General Research Protocols and Methods

Plasmid Isolation and Analysis. Pictorial Protocol. Mike Zeller. - [Read Plasmid Isolation and Analysis. Pictorial 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 DNA for Pulsed-field Gel Electrophoresis: Isolation of DNA from Mammalian Cells - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot4030

Category: Nucleic Acid Electrophoresis Protocols: Pulsed Field Gel Electrophoresis Protocols

Protocol for preparation of DNA for pulsed-field gel electrophoresis: isolation of DNA from mammalian cells and tissues. Genomic DNAs from mammalian cells are prepared for pulsed-field gel electrophoresis by lysing cells in situ in an agarose plug. Following digestion with an appropriate restriction enzyme, the plug is loaded directly into the well of a pulsed-field gel or it can be melted before loading. - [Read Preparation of DNA for Pulsed-field Gel Electrophoresis: Isolation of DNA from Mammalian Cells]

Preparation of DNA for Pulsed-field Gel Electrophoresis: Isolation of Intact DNA from Yeast - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3235

Category: Nucleic Acid Electrophoresis Protocols: Pulsed Field Gel Electrophoresis Protocols

Protocol for preparation of DNA for pulsed-field gel electrophoresis: isolation of intact DNA from yeast. Yeast cells are first treated enzymatically to break down the cell walls and then resuspended in low-melting-temperature agarose plugs. The DNA is liberated by infusing the plugs with lysis buffer and proteases. This method is used to prepare both conventional and artificial yeast chromosomes. - [Read Preparation of DNA for Pulsed-field Gel Electrophoresis: Isolation of Intact DNA from Yeast]

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]

Preparing Lambda DNA - http://sequence-www.stanford.edu/protocols/lambdaDNAprep.html

Category: Molecular Biology Protocols: Phage Protocols and Methods: Lambda Phage Protocols

Detailed Lambda DNA Isolation and Extraction protocol with recipes. Stanford DNA Sequence & Technology Center. - [Read Preparing Lambda DNA]

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 Isolation and Analysis From Plants - http://csm.jmu.edu/biology/courses/bio480_580/mblab/protein_isolation.htm

Category: Protein Protocols: Protein Extraction From Tissue

Protein Isolation and Analysis From Plants. T. Rife. Department of Biology, James Madison University. Harrisonburg, VA - [Read Protein Isolation and Analysis From Plants]

Protein isolation with tissue array construction - http://147.52.72.117/ijmm/2003/volume12/number3/299.pdf

Category: Protein Protocols: Protein Extraction From Tissue

Combination of nucleic acid and protein isolation with tissue array construction: Using defined histologic regions in single frozen tissue blocks for multiple research purposes - [Read Protein isolation with tissue array construction]

Protocol for Isolation, Cryopreservation, and Thawing of PBMCs - http://www.biomedcentral.com/content/supplementary/1471-2172-6-17-S1.pdf

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

Cryopreserved PBMCs are a common specimen source for studies of immunological responses to vaccines, immunotherapies, etc. The health and viability of cells recovered post-cryopreservation are of course critical to the success and accuracy of immunological assays performed on them. This protocol standardizes PBMC isolation and cryopreservation techniques, specifically for the assessment of thawed cells by cytokine flow cytometry. - [Read Protocol for Isolation, Cryopreservation, and Thawing of PBMCs]

Protocol for the Isolation of a Heterogenous Muscle-Derived Cell Population - http://www.bcm.edu/labs/goodell/protocols/Goodell_Musclestemcells.pdf

Category: Cell Organelle Protocols: Subcellular Fractionation Protocols

This protocol is employed for the purification of a population of muscle-derived cells from skeletal muscle of C57Bl/6 animals. The resulting preparation is a mixture of many cell types including satellite cells (a.k.a., muscle progenitor cells) and hematopoietically active muscle-derived cells. - [Read Protocol for the Isolation of a Heterogenous Muscle-Derived Cell Population]

Protocol for the Isolation of Pancreatic Duct Epithelial Cells - http://pen2.igc.gulbenkian.pt/cftr/vr/b/gray_protocol_for_the_isolation_of_pancreatic_duct.pdf

Category: Cell Biology and Cell Culture: Epithelial Cell Culture

Protocol for the Isolation of Pancreatic Duct Epithelial Cells. Gray and Argent. - [Read Protocol for the Isolation of Pancreatic Duct Epithelial Cells]

Protocol Guide for the N. crassa Yeast Artificial Chromosome Library - http://www.fgsc.net/fgn41/yac1.html

Category: Fungi Protocols: Neurospora Protocols

Protocol guide for the N. crassa yeast artificial chromosome library. Includes: Chromosome Walking; Hybridization screening of the YAC library; YAC restriction mapping and contig building; Preparation of chromosomal DNA plugs of YAC clones; Partial restriction enzyme digestion of YAC DNA plugs; Using CHEF gel analysis to resolve YAC clones; Southern Hybridization; Isolation of terminal restriction fragments from cloned DNA inserts in YAC clones; etc. - [Read Protocol Guide for the N. crassa Yeast Artificial Chromosome Library]

Protocols for Recombinant DNA Isolation, Cloning, and Sequencing. - http://www.genome.ou.edu/protocol_book/protocol_index.html

Category: DNA Protocols: DNA Cloning

Protocols for Recombinant DNA Isolation, Cloning, and Sequencing. Roe and Crabtree. - [Read Protocols for Recombinant DNA Isolation, Cloning, and Sequencing.]

Articles (1-5 of 5)

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.

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.

DNA Fragment Purification from Polyacrylamide Gels Protocol

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

DNA Ligation Protocol

The DNA Ligation protocol described here contains the steps required to join together using ligase enzyme both plasmid DNA and insert DNA fragments in order to create a new plasmid. This new ligated plasmid can be transformed after into competent bacteria to produce DNA for mini, midi or maxi-prep isolation.

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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