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Acrylamide Gels for Resolving Small (25bp-700bp) DNA Fragments Protocol - http://www.oardc.ohio-state.edu/stockingerlab/Protocols/AcrylamideGels%28DNA%29.pdf

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

Protocol for acrylamide gels for resolving small (25bp-700bp) DNA fragments. - [Read Acrylamide Gels for Resolving Small (25bp-700bp) DNA Fragments Protocol]

Addition of linkers/adaptors to blunt-end DNA fragments PDF - http://www.oup.co.uk/pdf/pas/9v1-7-11.pdf

Category: Molecular Biology Protocols: DNA Ligation Protocols

Addition of linkers/adaptors to blunt-end DNA fragments. Ligation of mixture containing DNA and linkers/adaptors. Powell Gannon. Oxford Practical Approach Series. - [Read Addition of linkers/adaptors to blunt-end DNA fragments PDF]

AFLP: a new technique for DNA fingerprinting. - http://www.pubmedcentral.gov/articlerender.fcgi?tool=pubmed&pubmedid=7501463

Category: PCR Protocols: AFLP PCR

P Vos, R Hogers, M Bleeker, M Reijans, T van de Lee, M Hornes, A Frijters, J Pot, J Peleman, and M Kuiper. 1995. Nucleic Acid Research. The AFLP technique is based on the selective PCR amplification of restriction fragments from a total digest of genomic - [Read AFLP: a new technique for DNA fingerprinting.]

Analysis of DNA Fragmentation Using Agarose Gel Electrophoresis (Subscription Required) - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4429

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

Analysis of DNA Fragmentation Using Agarose Gel Electrophoresis Shailaja Kasibhatla et al. This protocol provides a qualitative method for assessing cell death by detecting DNA fragments using agarose gel electrophoresis. One of the classic features of apoptosis is the cleavage of the genomic DNA into oligonucleosomal fragments represented by multiples of 180-200 bp. Visualizing these fragments can aid in characterizing an apoptotic event. May be combined with more quantitative methods. - [Read Analysis of DNA Fragmentation Using Agarose Gel Electrophoresis (Subscription Required)]

Analysis of DNA Fragmentation Using Propidium Iodide (PI) Fluorescence of Individual Nuclei Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4430

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols: DNA Staining Protocols for Flow Cytometry

This protocol provides a method for analysis of DNA fragmentation using flow cytometry after propidium iodide (PI) labeling of apoptotic nuclei. Apoptotic nuclei stain less than their normal counterparts because of diffusion of low-molecular-weight fragments out of the cells and/or chromatin condensation. - [Read Analysis of DNA Fragmentation Using Propidium Iodide (PI) Fluorescence of Individual Nuclei Protocol]

Analysis Of DNA Fragmentation Using The JAM Assay (Subscription Required) - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4432

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

Analysis of DNA Fragmentation Using the JAM Assay. By Shailaja Kasibhatla et al., The JAM assay is based on labeling nuclear DNA of cycling cells with [3H]thymidine and harvesting samples on glass fiber filters. Apoptosis will generate DNA fragments small enough to pass through the glass fiber filter, resulting in decreased radioactivity of the particular sample. Cell-mediated cytotoxicity or cell killing mediated by cytotoxic T lymphocytes (CTL) can also be measured by this technique. - [Read Analysis Of DNA Fragmentation Using The JAM Assay (Subscription Required)]

Bidirectional Cloning of PCR Products Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4139

Category: PCR Protocols: PCR Cloning Protocols

Protocol is for bidirectional, blunt-end cloning of DNA fragments. The target DNA is PCR amplified and 3'-extensions are polished with Pfu DNA polymerase. The amplicon is ligated to a blunt-ended plasmid DNA, and the products of the ligation reaction are used to transform competent Escherichia coli. A restriction enzyme is added to the ligation reaction to relinearize any self-religating vector DNA. - [Read Bidirectional Cloning of PCR Products Protocol]

Chromatin Immunoprecipitation (ChIP) of Protein Complexes Protocol - http://www.cshprotocols.org/cgi/content/abstract/2006/23/pdb.prot4560

Category: Immunology: Immunoprecipitation Protocols: Chromatin Immunoprecipitation Protocols

Protocol describes the use of chromatin immunoprecipitation technology (ChIP) to analyze interactions of proteins or protein complexes with DNA in vivo. In this approach, the material is fixed with formaldehyde to preserve DNA-protein and protein-protein associations, the cells are lysed, and the chromatin is cut and solubilized. The chromatin suspension is immunoprecipitated with an antibody against the protein(s) of interest, and the coimmunoprecipitated DNA fragments are analyzed. - [Read Chromatin Immunoprecipitation (ChIP) of Protein Complexes Protocol]

Chromatin Immunoprecipitation and Microarray-Based Analysis of Protein Location Protocol - http://www.nature.com/nprot/journal/v1/n2/abs/nprot.2006.98.html

Category: Immunology: Immunoprecipitation Protocols: Chromatin Immunoprecipitation Protocols

Genome-wide location analysis, also known as ChIP-Chip, combines chromatin immunoprecipitation and DNA microarray analysis to identify protein-DNA interactions that occur in living cells. Protein-DNA interactions are captured in vivo by chemical crosslinking. Cell lysis, DNA fragmentation and immunoaffinity purification of the desired protein will co-purify DNA fragments that are associated with that protein. - [Read Chromatin Immunoprecipitation and Microarray-Based Analysis of Protein Location Protocol]

Cleavage Efficiency Close to the Termini of PCR Fragments - http://www.fermentas.com/techinfo/re/restrdigpcrii.htm

Category: Molecular Biology Protocols: Restriction Enzyme Digestion

Cleavage Efficiency Close to the Termini of PCR Fragments. List of restriction enzymes and their cleavage efficiencies near the ends of DNA. Fermentas - [Read Cleavage Efficiency Close to the Termini of PCR Fragments]

Construction of Genomic DNA Libraries in Cosmid Vectors Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot4000

Category: DNA Protocols: Cosmid Library Protocols

Protocol describes how to construct a library of 35-45-kb fragments of genomic DNA in the double cos site cosmid vector, SuperCos-1. The steps include: Linearization and dephosphorylation of SuperCos-1 DNA; Partial digestion of high-molecular-weight DNA with MboI; Dephosphorylation of high-molecular-weight genomic DNA; Ligation of cosmid arms to genomic DNA: Packaging and plating recombinants; Isolation and analysis of recombinant cosmids: Validation of the library. - [Read Construction of Genomic DNA Libraries in Cosmid Vectors Protocol]

Direct Retrieval of DNA Fragments from Pulsed-field Gels Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot4083

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

Protocol for direct retrieval of DNA fragments from pulsed-field gels. A gel slice containing a fragment of DNA resolved by pulsed-field gel electrophoresis is treated with agarase. The released DNA can be used as a substrate for ligation or restriction without further purification. - [Read Direct Retrieval of DNA Fragments from Pulsed-field Gels Protocol]

Directional Cloning of PCR Products Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4140

Category: PCR Protocols: PCR Cloning Protocols

Protocol is for directional blunt-end cloning of DNA fragments. The target DNA is PCR-amplified, 3'-extensions are polished with Pfu DNA polymerase, and the amplicon is ligated to a blunt-ended plasmid DNA. The products of the ligation reaction are used to transform competent Escherichia coli. A restriction enzyme is added to the ligation reaction to relinearize any self-religating vector DNA. - [Read Directional Cloning of PCR Products Protocol]

DNA Fragment Purification from Agarose or Acrylamide - http://axon.med.harvard.edu/~cepko/protocol/mike/D5.html

Category: DNA Protocols: DNA Extraction Protocols: DNA Extraction Polyacrylamide Gels

DNA Fragment Purification from Agarose or Acrylamide. The protocol for fragments from 200 bp to 10 kb the agarose purification is ideal. For smaller fragments (20 bp to 400 bp) the acrylamide purification is preferred. - [Read DNA Fragment Purification from Agarose or Acrylamide]

DNA Fragment Purification from Agarose or Acrylamide Protocol - http://genetics.med.harvard.edu/~cepko/protocol/mike/D5.html

Category: Nucleic Acid Purification Protocols: DNA Isolation Protocols

Protocol for DNA fragment purification from agarose or acrylamide. For fragments from 200 bp to 10 kb the agarose purification is ideal. For smaller fragments (20 bp to 400 bp) the acrylamide purification is preferred. - [Read DNA Fragment Purification from Agarose or Acrylamide Protocol]

DNA Fragment Purification from Agarose Protocol - http://axon.med.harvard.edu/~cepko/protocol/mike/D5.html

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

DNA Fragment Purification from Agarose Protocol. This protocol is best for fragments from 200 bp to 10 kb the agarose purification is ideal. For smaller fragments (20 bp to 400 bp) the acrylamide purification is preferred. - [Read DNA Fragment Purification from Agarose Protocol]

DNA Ligation - http://www.bio.com/protocolstools/protocol.jhtml?id=p2053

Category: Molecular Biology Protocols: DNA Ligation Protocols

Description and protocol of the steps required to join together plasmid and insert fragments to create a new plasmid. Paul Kaufman, Univ California. - [Read DNA Ligation]

EMBL Cloning protocols - http://www.embl-hamburg.de/display?file=~geerlof/webPP/protocoldb/Cloning/pdb_cloning.html

Category: DNA Protocols: DNA Cloning

Preparation of oligo solutions PCR experiments, Digestion of insert DNA, Digestion and dephosphorylation of vector DNA, Ligation of DNA fragments with sticky ends, Ligation of DNA fragments with blunt ends, Preparation of chemically competent E. coli - [Read EMBL Cloning protocols]

Fill-in Labeling of DNA Fragments Protocol - http://genetics.med.harvard.edu/~cepko/protocol/mike/F1.html

Category: Nucleic Acid Modification Protocols: Nucleic Acid Labeling Protocols

Protocol for fill-in labeling of DNA fragments. - [Read Fill-in Labeling of DNA Fragments Protocol]

Fluorometric Quantitation of DNA Using Hoechst 33258 Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4458

Category: Nucleic Acid Detection Protocols

Protocol describes the quantitation of DNA using Hoechst 33258, a fluorescent dye that binds to double-stranded DNA. Fluorometry is simple and more sensitive than spectrophotometry, and allows the detection of nanogram quantities of DNA. The assay can only be used to measure the concentration of DNAs whose sizes exceed ~1 kb, as Hoechst 33258 binds poorly to smaller DNA fragments. - [Read Fluorometric Quantitation of DNA Using Hoechst 33258 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]

Germ Layer Explant Recombination Culture Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4369

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

This protocol describes a method for recombining and culturing germ layer fragments. It is useful for testing the inductive properties of fragments from wild-type and mutant mouse embryos. - [Read Germ Layer Explant Recombination Culture Protocol]

Germ Layer Explant Recombination Culture Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4369

Category: Mouse Protocols: Mouse Cell Culture Protocols

Protocol describes a method for recombining and culturing germ layer fragments. It is useful for testing the inductive properties of fragments from wild-type and mutant mouse embryos. - [Read Germ Layer Explant Recombination Culture Protocol]

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]

How to Separate Chromosomes and Restriction Fragments Using Pulsed Field Gel Electrophoresis - http://www.fgsc.net/neurosporaprotocols/How%20to%20separate%20chromosomes.pdf

Category: Fungi Protocols: Neurospora Protocols

Information and tips on how to separate chromosomes and restriction fragments using pulsed field gel electrophoresis. - [Read How to Separate Chromosomes and Restriction Fragments Using Pulsed Field Gel Electrophoresis]

Articles (1-6 of 6)

Antibodies Phage Expression Protocol Using 96-well Microtiter Plates

A Phage display expression protocol for antibody expression in 96-well microtiter plates.

Purifying DNA Fragments from Agarose Gels Alternate Protocol

An alternate protocol for purifying DNA fragments from agarose gels.

DNA Fragment Purification from Polyacrylamide Gels Protocol

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

Quick DNA Agarose Gel Electrophoresis Protocol.

A quick and simple protocol for the electrophoresis of DNA fragments using agarose 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.

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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