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"CORE SAMPLE" PCR - A method to re-PCR unique bands from products of mixed size - http://www.mcb.uct.ac.za//corepcr.htm

Category: DNA Protocols: PCR Polymerase Chain Reaction Protocols: Standard PCR Protocol

"CORE SAMPLE" PCR - A method to re-PCR unique bands from products of mixed size - Molecular Biology Techniques Manual - Rybicki - [Read "CORE SAMPLE" PCR - A method to re-PCR unique bands from products of mixed size]

"CORE SAMPLE" PCR - A method to re-PCR unique bands from products of mixed size - http://www.mcb.uct.ac.za//corepcr.htm

Category: PCR Protocols: Standard PCR Protocol

5' RACE Protocol For Generation of Seq. Tags - http://baygenomics.ucsf.edu/protocols/comp1/RACE_generation.pdf

Category: RNA Protocols: 5' RACE - Rapid Amplification of cDNA Ends Protocols

Can also use 96-well plates. First Strand cDNA Synthesis, First Size selection, second 2nd strand cDNA synthesis. Bay Area Functional Genomics Consortium, UCSF. - [Read 5' RACE Protocol For Generation of Seq. Tags]

Advanced Methodologies in Flow Cytometry - http://www.cyto.purdue.edu/flowcyt/research/cytotech/amfc/data/index1.htm

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

Advanced Methodologies in Flow Cytometry. Andrea Cossarizza et al.. Materials from a workshop held March, 1997 in Modena, Italy. - [Read Advanced Methodologies in Flow Cytometry]

Agarose Gel Preparation Protocol - http://fp.bio.utk.edu/mycology/Techniques/mt-agarose_gels.htm

Category: Molecular Biology Protocols: Agarose Gel Preparation

Solutions for Agarose gel prepartion, Selection of Appropriate Gel Size and Composition. Mycology Lab University of Tennessee - [Read Agarose Gel Preparation Protocol]

Alkaline Agarose Gel Electrophoresis Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot4027

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

Alkaline agarose gels are used chiefly to measure the size of first and second strands of cDNA (Construction of cDNA Libraries Stage 1: Synthesis of First-strand cDNA Catalyzed by Reverse Transcriptase) and to analyze the size of the DNA strand after digestion of DNA-RNA hybrids with nucleases such as S1. - [Read Alkaline Agarose Gel Electrophoresis Protocol]

Analysis of Recombinant Bacteriophage M13 Clones Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3996

Category: Viral Manipulation Protocols: M13 Phage Manipulation Protocols

A rapid method to analyze the size of the single-stranded DNA of M13 recombinants. - [Read Analysis of Recombinant Bacteriophage M13 Clones Protocol]

Ancient DNA: Extraction, characterization, molecular cloning, and enzymatic amplification. PDF - http://www.pnas.org/cgi/reprint/86/6/1939

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

The DNA obtained was invariably of a low average molecular size and damaged by oxidative processes, but PCR can be used to amplify and study short mitochondrial DNA sequences thatare of anthropological and evolutionary significance. SVANTE PAABO, PNAS. - [Read Ancient DNA: Extraction, characterization, molecular cloning, and enzymatic amplification. PDF]

Apoptosis Pathway Poster Caspases - http://www.upstate.com/img/pathways/apoptosis_2003.jpg

Category: Cell Biology and Cell Culture: Apoptosis Protocols: Apoptosis Information

Apoptosis Pathway Mid-size Poster. Upstate. - [Read Apoptosis Pathway Poster Caspases]

C25 Isolation from spleen, thymus, pancreas, alveolar tissue and other tissues - http://www.axis-shield.com/densityhome/optiprep/C25.pdf

Category: Cell Biology and Cell Culture

OptiPrep Application sheet C14 describes procedures for determining the density and sedimenting properties of any cell (of any size or density) using either a continuous or discontinuous gradient of iodixanol. This Application Sheet describes procedures aimed at isolating specifically a relatively low-density cell fraction from any tissue. - [Read C25 Isolation from spleen, thymus, pancreas, alveolar tissue and other tissues]

Cell Fractionation of a Mixed Population of Cells - http://www.axis-shield.com/densityhome/optiprep/C14.pdf

Category: Cell Biology and Cell Culture

Cell fractionation techniques are presented for the design of gradient systems for separating one or more cell types from lavages of body cavities or from mechanically or enzymically-dissociated tissues. Includes: Preparation of cell suspension for gradient loading; Fractionation by buoyant density; Fractionation on the basis of cell size. - [Read Cell Fractionation of a Mixed Population of Cells]

Cell Synchronization Using Centrifugal Elutriation Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/18/pdb.prot4490

Category: Cell Biology and Cell Culture: Cell Cycle Protocols

protocol describes a method for the synchronization of cell populations using centrifugal elutriation. The method relies on the fact that cell size increases linearly as cells proceed through the cell cycle. Cells of similar size (and cell cycle phase) are eluted stepwise from the cell chamber, with the smallest size (those in early G1) being eluted first. Using this procedure, it is possible to obtain relatively pure populations of cells at various points in G1, S, and G2/M. - [Read Cell Synchronization Using Centrifugal Elutriation Protocol]

Cell Synchronization Using Centrifugal Elutriation Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/18/pdb.prot4490

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

This protocol describes a method for the synchronization of cell populations using centrifugal elutriation. The method relies on the fact that cell size increases linearly as cells proceed through the cell cycle. Cells of similar size (and cell cycle phase) are eluted stepwise from the cell chamber, with the smallest size (those in early G1) being eluted first. Using this procedure, it is possible to obtain relatively pure populations of cells at various points in G1, S, and G2/M. - [Read Cell Synchronization Using Centrifugal Elutriation Protocol]

Column Chromatography Protocol - http://www.biochem.arizona.edu/classes/bioc463a/Info/protocols/exp_4_1.pdf

Category: Chromatography Protocols: Column Chromatography Protocols

Size Exclusion Column Chromatography Protocol. PDF. In this protocol you will learn how to use three types of column chromatography: Gel Filtration or Size Exclusion (SEC), Ion Exchange (IEC), and affinity (AC) in order to purify proteins and enzymes based on the physical properties of these biomolecules. Univ. Arizona, Biochemistry. - [Read Column Chromatography 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]

Differentiation of Embryonic Stem (ES) Cells Using the Hanging Drop Method - http://www.cshprotocols.org/cgi/content/extract/2006/18/pdb.prot4485

Category: Stem Cell Protocols: Stem Cell Culture Protocols

In vitro differentiation of ES cells occurs when the cells are allowed to aggregate in suspension culture in the absence of mouse embryonic fibroblast (MEF) feeders and leukemia inhibitory factor (LIF). Hanging drops provide a uniform aggregate size, which is then expanded by continued growth in suspension culture. The embryoid bodies are then plated and allowed to differentiate further in culture. - [Read Differentiation of Embryonic Stem (ES) Cells Using the Hanging Drop Method]

Dye Protocols and Notes for Cosmid, BAC, BAC, Fosmid Templates - http://www.genome.ou.edu/big_dyes_cos_bac_pac_fos.html

Category: BACs Bacterial Artificial Chromosome Protocols

Sequencing conditions tested for the ABI Big-Dye terminators (PE-ABI #4303150 for the 1000 reaction kit - Description: TF,KIT BTD RR-1000) with various templates. Important to quantitate all templates by agarose gel electrophoresis vs size and concentration standards and do a few tests with different template concentrations to determine the optimal conditions for your reactions. Several conditions are given. - [Read Dye Protocols and Notes for Cosmid, BAC, BAC, Fosmid Templates]

Early Events in B Lymphocyte Activation Protocol - https://catalog.invitrogen.com/index.cfm?fuseaction=iProtocol.unitSectionTree&treeNodeID=9E6634190D367003ECF94AE65295FA77&objectid=6674AB8295693316922D8DFC029193A6

Category: Immunology: B Cell Protocols

B cell activation can be quantitated indirectly by assaying antibody production or directly by measuring cellular changes that occur immediately after exposure to an activation signal. Provides methods for the latter (direct) approach--namely, methods for quantifying early parameters of B cell activation such as increases in intracellular ionized calcium concentration [Ca2+]I, cell size, and MHC class II-antigen expression. - [Read Early Events in B Lymphocyte Activation Protocol]

Electroporation for Sindbis Virus Production Protocol - http://bio.research.ucsc.edu/people/hinck/electroporation.html

Category: Transfection Protocols: Electroporation Sonoporation Protocols

Protocol for electroporation for Sindbis virus production. - [Read Electroporation for Sindbis Virus Production Protocol]

Ethics, Guidelines for Animal Research PDF - http://www.brl.uic.edu/courses/TNotes05.pdf

Category: Animal Research Techniques

Needs of animals, Ventilation, lighting and temperature considerations. Cage size/density. Handling. Blood Collection and guidlines. EXCELLENT GUIDE GC470 ESSENTIALS FOR ANIMAL RESEARCH BRL Univ. Illinois. - [Read Ethics, Guidelines for Animal Research PDF]

Extraction of Total Protein from Arabidopsis Protocol - http://www.cshprotocols.org/cgi/content/abstract/2007/1/pdb.prot4680

Category: Plant Biology Protocols: Arabidopsis Protein Peptide Protocols

The simplest way to analyze proteins is in unfractionated extracts. However, it is often desirable to fractionate proteins, e.g, by size. This procedure extracts total protein from Arabidopsis samples. Typical yields are ~2-3 mg/ml (using rosette leaves) or 6-8 mg/ml (using young seedlings). - [Read Extraction of Total Protein from Arabidopsis Protocol]

Fragmentation of DNA by Nebulization Protocol - http://www.cshprotocols.org/cgi/content/abstract/2006/23/pdb.prot4539

Category: Cloning Protocols

Protocol describes a method for DNA fragmentation by nebulization, in which the fine mist created by forcing a DNA solution through a small hole in the nebulizer unit is collected. The size of the fragments obtained by nebulization is determined chiefly by the speed at which the DNA solution passes through the hole, altering the pressure of the gas blowing through the nebulizer, the viscosity of the solution, and the temperature. - [Read Fragmentation of DNA by Nebulization Protocol]

Generation of Bidirectional Sets of Deletion Mutants by Digestion with BAL 31 Nuclease Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3589

Category: Genetics and Genomics: Genotyping Protocols: Mutation Detection Protocols

In this method, the nuclease BAL 31 is used to make uni- or bidirectional deletions in a segment of cloned DNA. BAL 31 is a complex enzyme and tends to digest a population of double-stranded DNA targets in an asynchronous fashion, Deletions created by BAL 31 are therefore far more heterogeneous in size than those created by processive enzymes such as exonuclease III. - [Read Generation of Bidirectional Sets of Deletion Mutants by Digestion with BAL 31 Nuclease Protocol]

Isolation of DNA Fragments from Polyacrylamide Gels by the Crush and Soak Method Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot2936

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

The "crush and soak" method, which works best for DNAs <1 kb in size, can be used to recover both singleand double-stranded DNAs from polyacrylamide gels. The yield of eluted DNA varies from <30% to >90% depending on the size of the DNA fragment. - [Read Isolation of DNA Fragments from Polyacrylamide Gels by the Crush and Soak Method Protocol]

Isolation of Ly-1+/CD5+ B Cells by Cell Sorting Protocol - https://catalog.invitrogen.com/index.cfm?fuseaction=iProtocol.unitSectionTree&treeNodeID=9E66368A929753C320BDC7C58DA3FDFD&objectid=66749F0EB1D3C751C86C30427F3C7AB0

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

A flow cytometry technique is presented, which results in the selection and isolation of two populations of cells from a complex mixture based on physical properties (e.g., size and internal granularity) and correlated expression of several surface molecules - [Read Isolation of Ly-1+/CD5+ B Cells by Cell Sorting Protocol]

Articles (1-5 of 5)

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.

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.

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.

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.

Vector Design Protocol for Transgenic Mice Generation

The protocol gives general considerations for the design of targeting vectors for transgenic mice. The protocol shares tips in the design of knock-out and knock-in vectors and some of their strategies for producing homologously recombined embryonic stem cells.

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