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DNA Microprep Isolation from Plants Protocol - http://www.bio.com/protocolstools/protocol.jhtml?id=p9010

Category: Plant Biology Protocols: Plant DNA RNA Isolation Protocols

The original maize DNA miniprep protocol is used extensively for many plant species and different tissues. This slightly modified version is acceptable for most DNA extractions. The procedure has the advantage of isolating DNA from plant material very rapidly. The procedure requires a table-top drill-press (mechanized homogenizer). - [Read DNA Microprep Isolation from Plants Protocol]

DNA Miniprep Isolation from Plants Protocol - http://www.bio.com/protocolstools/protocol.jhtml?id=p1079

Category: Plant Biology Protocols: Plant DNA RNA Isolation Protocols

The original maize DNA miniprep protocol is used extensively for many plant species and different tissues. This slightly modified version is acceptable for most DNA extractions. The procedure has the advantage of speed and its use of inexpensive reagents. - [Read DNA Miniprep Isolation from Plants Protocol]

DNA-Binding Protein Purification with Dynabeads. - http://www.pubmedcentral.nih.gov/articlerender.fcgi?tool=pubmed&pubmedid=2671937

Category: DNA Protocols: EMSA DNA-Protein Protocols

Magnetic DNA affinity purification of yeast transcription factor tau--a new purification principle for the ultrarapid isolation of near homogeneous factor. Gabrielsen et al. 1989 - [Read DNA-Binding Protein Purification with Dynabeads.]

Electroporation and isolation of ES cell lines Mouse - http://baygenomics.ucsf.edu/protocols/comp1/electroporation.html

Category: Cell Biology and Cell Culture: ES Stem Cell Biology and Culture

Electroporation and isolation of ES cell lines Mouse. BayGenomics. - [Read Electroporation and isolation of ES cell lines Mouse]

Fractionation of T and B Cells Using Magnetic Beads Protocol - https://catalog.invitrogen.com/index.cfm?fuseaction=iProtocol.unitSectionTree&treeNodeID=9E66367B022BC1A53B67DF9C702B2513&objectid=66749A4CD26365F08DFBDE734F6A50F5

Category: Immunology: Mononuclear Cell Isolation Protocols: B Lymphocyte Isolation Protocols

Protocol describes the purification of mouse T cells, B cells, and T cell subsets using magnetic bead separation. Isolation of cell subsets using magnetic beads is quick, simple, and reliable and can result in high yields of very pure cells. - [Read Fractionation of T and B Cells Using Magnetic Beads Protocol]

Category: Immunology: Mononuclear Cell Isolation Protocols: T-Cell Isolation Protocols

Fractionation of Vacuolar and Subvacuolar vesicles and Vacuole and Cytoplasm-to-Vacuole Targeting - http://www.axis-shield.com/densityhome/optiprep/S29.pdf

Category: Cell Organelle Protocols: Subcellular Fractionation Protocols

Fractionation of (a) vacuolar and subvacuolar vesicles and (b) vacuole and cytoplasm-to-vacuole targeting (Cvt) vesicles from yeast spheroplasts in a pre-formed discontinuous iodixanol gradients. Protocol includes: Formation of yeast spheroplasts; Isolation and vesiculation of the vacuoles; Separation of the vacuolar and subvacuolar vesicles; Separation of vacuoles and Cvt vesicles from a yeast spheroplast lysate. - [Read Fractionation of Vacuolar and Subvacuolar vesicles and Vacuole and Cytoplasm-to-Vacuole Targeting]

Fungal DNA Isolation Method - http://wheat.pw.usda.gov/~lazo/methods/lazo/dnafun.html

Category: DNA Protocols: DNA Extraction Protocols

DNA isolation from various fungal species including: Cochliobolus, Aternaria, and Fusarium. Key steps: (1) the use of young lyophilized mycelial mats - yield less contaminating carbohydrates; (2) proteinase K in the extraction buffer to destroy DNases (f - [Read Fungal DNA Isolation Method]

Fungal DNA Isolation Protocol - http://wheat.pw.usda.gov/~lazo/methods/lazo/dnafun.html

Category: Nucleic Acid Purification Protocols: DNA Isolation Protocols

Protocol for fungal DNA isolation. The key elements in this prep are (1) the use of young lyophilized mycelial mats....young mats (4 days growth for C. carbonum)...yield less contaminating carbohydrates and other misc. junk (2) lots of proteinase K in the extraction buffer to kill Dnases (final =0.3mg/ml). - [Read Fungal DNA Isolation Protocol]

Gene Expression and Protein Localization in Arabidopsis Protocols - http://www.arabidopsis.org/cshl-course/7-gene_expression.html

Category: Plant Biology Protocols: Arabidopsis Genetics Protocols

Protocols for gene expression and protein localization in Arabidopsis. Includes: Detection of the native protein; Detection of a recombinant version; Immunofluorescence detection in Arabidopsis protoplasts; Isolation of Arabidopsis seedling protoplasts; Subcellular localization of GUS-fusion proteins in Arabidopsis seedlings; Localization of Arabidopsis proteins with GUS in situ enzyme assay. - [Read Gene Expression and Protein Localization in Arabidopsis Protocols]

Genomic DNA isolation from blood - http://www.genome.ou.edu/protocol_book/protocol_partIII.html#III.H

Category: DNA Protocols: DNA Extraction Blood

Genomic DNA isolation from blood. Roe Lab - [Read Genomic DNA isolation from blood]

Getting DNA from Old Dead Stuff - http://www.uga.edu/srel/DNA_Lab/OldDNA99.rtf

Category: DNA Protocols: DNA Extraction Protocols: Extracting Ancient DNA

DNA Isolation from museum specimens: 1) what's the best tissue to sample; 2) what's the best way to get DNA out of that tissue? 1) the easiest well-dried tissue to get, and 2) the easiest extraction methods you are comfortable doing. Travis Glenn. - [Read Getting DNA from Old Dead Stuff]

Glass Milk Agarose Gel DNA Extracton Protocol - http://mycoplasmas.vm.iastate.edu/lab_site/methods/DNA/glass_milk.html

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

Isolation of DNA fragments using glass milk (GENE-CLEAN). Glass Milk Agarose Gel DNA Extracton Protocol. Minion Lab, College of Veterinary Medicine at Iowa State University. - [Read Glass Milk Agarose Gel DNA Extracton Protocol]

Growing Bacteriophage M13 in Liquid Culture Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3994

Category: Viral Culture and Isolation Protocols: Viral Culture Protocols

Most manipulations with M13, including preparations of viral stocks and isolation of single- and double-stranded DNAs, begin with small-scale liquid cultures that are infected with an M13 plaque, picked from an agar plate. - [Read Growing Bacteriophage M13 in Liquid Culture Protocol]

Growth of Primary Cell Culture and Viral Vector Handling Protocols - http://www.stanford.edu/dept/EHS/prod/researchlab/bio/docs/Palmer_Bio_protocol.pdf

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

The procedures involve the isolation and growth of primary cell cultures from rodent and human tissue as well as the use of viral vectors for the introduction and expression of mammalian genes in cells in culture and in live rodents. - [Read Growth of Primary Cell Culture and Viral Vector Handling Protocols]

Growth of S. cerevisiae and Preparation of DNA Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3298

Category: Yeast Protocols: Yeast Genetics Protocols: YACs Protocols

Protocol describes methods for isolation of DNA from a strain of S. cerevisiae carrying a recombinant YAC. Because the linear YAC DNAs are sensitive to shearing forces, pipettes with wide-bore tips should be used to transfer DNAs. The method is suitable for preparing DNA that will be used for agarose gel electrophoresis, Southern blotting, subcloning, genomic library construction, PCR, or other methods that do not require intact high-molecular-weight DNA. - [Read Growth of S. cerevisiae and Preparation of DNA Protocol]

GUS Reporter Gene Assay Protocol - http://www.oardc.ohio-state.edu/stockingerlab/Protocols/GUS%20Assays.pdf

Category: Reporter Gene Assays

Protocol for GUS reporter gene assay. Includes: Protein isolation; Alternative method for small (<1g) quantities of tissue; GUS assays; Bradford protein concentration determination assays - [Read GUS Reporter Gene Assay Protocol]

Human Embryonic Stem Cell Protocols - http://stemcells.nih.gov/staticresources/research/protocols/BresaGen_hESC_manual_2.1.pdf

Category: Stem Cell Protocols: Stem Cell Culture Protocols

Human embryonic stem cells (hESCs) are pluripotent cells capable of differentiation to representatives of all three germ layers. Includes: Isolation of Primary Mouse Embryo Fibroblasts; Thawing and preparing p1 MEF feeder plates; Preparation of MEF- Conditioned Medium (MEF-CM; Microdissection Passaging of hESCs; Bulk passaging of hESC; Cryopreservation of hESCs; Thawing of hESCs; Karyotyping. - [Read Human Embryonic Stem Cell Protocols]

In Vitro Transcription Assay (Run-off Assay) using Permeabilized Cells - http://www.bio.com/protocolstools/protocol.jhtml?id=p1971

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

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]

Isolating BAC DNA from Bacterial Cultures Using the NucleoBond System Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/16/pdb.prot4390

Category: BACs Bacterial Artificial Chromosome Protocols

Protocol describes isolation of BAC DNA from bacterial cultures using the commercial NucleoBond system. - [Read Isolating BAC DNA from Bacterial Cultures Using the NucleoBond System Protocol]

Isolating Germ Cells from the Genital Ridge - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4370

Category: Mouse Protocols: Mouse Cell Culture Protocols

Protocol describes isolation of germ cells from the genital ridge of fetal mice from 11.5 days post coitum (dpc) onward. The germ cells can then be used for analysis, culture, or transplantation. - [Read Isolating Germ Cells from the Genital Ridge]

Isolating Germ Cells from the Genital Ridge Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4370

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

This protocol describes isolation of germ cells from the genital ridge of fetal mice from 11.5 days post coitum (dpc) onward. The germ cells can then be used for analysis, culture, or transplantation. - [Read Isolating Germ Cells from the Genital Ridge Protocol]

Isolating Mice Extraembryonic Membranes Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4367

Category: Mouse Protocols: Mouse Embryology Protocols

Isolation of extraembryonic tissues allows one to study normal mouse development as well as the molecular basis of defects which cause fetal death. This protocol describes a method for isolating extraembryonic membranes from pregnant mice. - [Read Isolating Mice Extraembryonic Membranes Protocol]

Isolating Postimplantation Embryos: Early Neural-Fold-Stage Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4365

Category: Stem Cell Protocols: Stem Cell Isolation Protocols

Isolation of postimplantation-stage embryos allows one to study normal development as well as genetic mutations which cause postimplantation defects. This protocol describes a method for isolation of early neural-fold-stage embryos. - [Read Isolating Postimplantation Embryos: Early Neural-Fold-Stage Protocol]

Isolating Postimplantation Embryos: Early Primitive-Streak-Stage Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4363

Category: Stem Cell Protocols: Stem Cell Isolation Protocols

Isolation of postimplantation-stage embryos allows one to study normal development as well as genetic mutations which cause post-implantation defects. This protocol describes a method for isolating early primitive-streak-stage embryos. - [Read Isolating Postimplantation Embryos: Early Primitive-Streak-Stage Protocol]

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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