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4% Paraformaldehyde Protocol - http://www.nottingham.ac.uk/pathology/protocols/paraform.html

Category: Immunohistochemistry Protocols: Fixation Protocols

Protocol for 4% paraformaldehyde. Includes: 4% PARAFORMALDEHYDE/PBS; 0.4% PARAFORMALDEHYDE/PBS (FOR POST-DIGESTION FIXATION). - [Read 4% Paraformaldehyde Protocol]

AFLP Method - http://haveylab.hort.wisc.edu/protocol/AFLP.html

Category: PCR Protocols: AFLP PCR

AFLP Procecure, includes restriction Digestion of Genomic DNA. Michael Havey - Vegetable Genetics Laboratory - [Read AFLP Method]

AFLP PCR Protocol PDF - http://www4.ncsu.edu/~wagebrey/AFLP-Salm.pdf

Category: PCR Protocols: AFLP PCR

Restriction digestion, Adapter ligation, Preamplification, Final Amplification steps for AFLP PCR. Detailed protocol. Wondwossen Abebe Gebreyes - [Read AFLP PCR Protocol PDF]

AFLP protocol - http://www.msu.edu/user/hazensam/aflp/AFLPprotocolMSU.html

Category: PCR Protocols: AFLP PCR

Restriction Digestion of Genomic DNA , Adapter preparation, Ligation of Adapters. Pre amplification Reactions. Gel electrophoresis, Silver Staining. AFLP Protocol and procedure. - [Read AFLP 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]

AR Protease Digestion Method - http://www.chemicon.com/techsupp/Protocol/Protease.asp

Category: Immunology: Antigen Retrieval

Protease Antigen Retrieval (Trypsin) Protocol. Chemicon. - [Read AR Protease Digestion Method]

Attachment of Linkers or Adaptors for Construction of cDNA Libraries - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3310

Category: DNA Protocols: cDNA Library Protocols

This stage achieves four goals: polishing the ends of double-stranded DNA, ligation of synthetic linkers or adaptors, digestion of the attached linkers to create cohesive termini, and preparing the cDNA for cloning. - [Read Attachment of Linkers or Adaptors for Construction of cDNA Libraries]

Bacteriophage Lamda Plant Genomic Library Construction Protocol - http://www.oardc.ohio-state.edu/stockingerlab/Protocols/GenomicDNAPhageLibraryConstruction.pdf

Category: DNA Protocols: Genomic DNA Library Protocols

Protocol for bacteriophage lamda plant genomic library construction. Includes: Preparation of the insert DNA; Pilot Sau3A Digestion; Full-scale partial digestions. - [Read Bacteriophage Lamda Plant Genomic Library Construction Protocol]

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]

DNA digestion protocol & hints PDF - http://www.crypto.wustl.edu/Protocols%20PDFs/RE%20digests%20(HL).pdf

Category: Molecular Biology Protocols: Restriction Enzyme Digestion

DNA digestion protocol & hints. Procedure and tips for restriction digestion. Doering laboratory, Department Molecular Microbiology, Washington University Medical School. - [Read DNA digestion protocol & hints PDF]

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]

Establishment of Stable Transfectant of CHO Lec Cells Protocol - http://www.cbrinstitute.org/labs/springer/protocols/jun_CHOlec.html

Category: Transfection Protocols: Stable Transfection Protocols

CHO Lec3.2.8.1 cells have 4 independent mutations in the N and O glycosylation pathways. When cultured with alpha-glucosidase I inhibitor N-butyl-deoxynojirimycin, glycoproteins produced in CHO Lec3.2.8.1 cells are completely susceptible to Endo H digestion. Endo H cleaves chitobiose, leaving a single N-linked N-acetylglucosamine per site which is ideal for maintenance of protein solubility and special carb-protein interactions, such as between the first N-acetyl glucosamine residue and tryp. - [Read Establishment of Stable Transfectant of CHO Lec Cells Protocol]

Factors that Influence Restriction Enzyme Digestion Activity - http://arbl.cvmbs.colostate.edu/hbooks/genetics/biotech/enzymes/cuteffects.html

Category: Molecular Biology Protocols: Restriction Enzyme Digestion

Factors that Influence Restriction Enzyme Digestion Activity. Buffer Composition, Incubation Temperature, DNA Methylation, Star Activity, Multiple Enzymes. Laura Austgen PhD et. al, Biomedical Hypertextbook, Colorado State Univ. - [Read Factors that Influence Restriction Enzyme Digestion Activity]

FIXATION PROTOCOL FOR FLOW CYTOMETRIC ANALYSIS OF DNA BY PI STAINING - http://www.vetmed.auburn.edu/index.pl/cell_cycle/dna_content

Category: Histology Protocols: Fixation of Cells Tissues Protocol

Analysis of Cell Cycle Based on DNA Content. FIXATION PROTOCOL FOR FLOW CYTOMETRIC ... Liberate cells from tissue culture flask by trypsin digestion and ... FIXATION PROTOCOL FOR FLOW CYTOMETRIC ANALYSIS OF DNA BY PI STAINING. Auburn University, College of Veterinary Medicine. You. - [Read FIXATION PROTOCOL FOR FLOW CYTOMETRIC ANALYSIS OF DNA BY PI STAINING]

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]

In Gel Digestion Protocols for Silver Stained and Coomassie Stained Gels - http://www.proteomics.com.cn/2005/papers/InGel.html

Category: Proteomic Protocols

In Gel Digestion Protocols for Silver Stained and Coomassie Stained Gels. (EMBL protocol). - [Read In Gel Digestion Protocols for Silver Stained and Coomassie Stained Gels]

In-gel digestion of proteins for peptide fingerprint mapping - http://www.bio.vu.nl/vakgroepen/mnb/proteomics/6.in-geldigestion.html

Category: Mass Spectrometry Protocols

Peptide Mapping Analysis Techniques. Dr.Aebersold, Vrije Universiteit Amsterdam - [Read In-gel digestion of proteins for peptide fingerprint mapping]

In-Gel Digestion of Proteins Separated by Polyacrylamide Gel Electrophoresis - http://web.archive.org/web/20010906052645/http://www.pharma.ethz.ch/bmm/protocols/ingeldig.html

Category: Proteomic Protocols: 2-D Gel Electrophoresis

In gel trypsin digestion for Mass spectrometry. (According to EMBL Protein and Peptide Group, 1997) - [Read In-Gel Digestion of Proteins Separated by Polyacrylamide Gel Electrophoresis]

In-gel digestion of proteins to be analyzed by mass spectrometry - http://www.narrador.embl-heidelberg.de/GroupPages/PageLink/activities/protocols/ingeldigest.html

Category: Proteomic Protocols

In-Gel Digestion Protocol. Excision of protein bands (spots) from polyacrylamide gels Reduction and alkylation . Reduction and alkylation . MALDI analysis of the supernatant after in-gel digestion. Extraction of Peptides.Matthias Wilm EMBL Bioanalytical R - [Read In-gel digestion of proteins to be analyzed by mass spectrometry]

In-gel Digestion protocol - http://www.hmc.psu.edu/core/oldsite/Maldi/malditofprotocols.htm

Category: Proteomic Protocols

In-gel Digestion protocol. BA Stanley. - [Read In-gel Digestion protocol]

Inverse PCR Protocol II - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3487

Category: PCR Protocols: Inverse PCR Protocols

Inverse PCR is used to amplify and clone unknown DNA that flanks one end of a known DNA sequence and for which no primers are available. The technique involves digestion by a restriction enzyme of a preparation of DNA containing the known sequence and its flanking region. The individual restriction fragments (many thousands in the case of total mammalian genomic DNA) are converted into circles by intramolecular ligation, and the circularized DNA is then used as a template in the PCR. - [Read Inverse PCR Protocol II]

Isolation of Adult Mouse Cardiac Myocytes from Two or More Hearts Protocol - http://www.signaling-gateway.org/data/cgi-bin/ProtocolFile.cgi/afcs_PP00000126.pdf?pid=PP00000126

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

This procedure describes the isolation and culture of adult mouse cardiac myocytes from two or more hearts. Includes modifications for the digestion of two or more hearts in the same procedure and subsequent pooling of myocytes derived from the multiple hearts. The isolation procedure is performed by one or more technicians and routinely yields approximately 1 million rod-shaped myocytes per heart. - [Read Isolation of Adult Mouse Cardiac Myocytes from Two or More Hearts Protocol]

Isolation of Dendritic Cells Protocol - https://catalog.invitrogen.com/index.cfm?fuseaction=iProtocol.unitSectionTree&treeNodeID=9E66369ADF1DF1962F01D3DA073183AB&objectid=6674A1FCBE1A61EE40A5E3EE8FB368F5

Category: Immunology: Mononuclear Cell Isolation Protocols

Presents two methods for preparing dendritic cells (DCs), a highly specialized type of antigen-presenting cell (APC). The first method involves the isolation of DCs from mouse spleen, resulting in a cell population that is highly enriched in accessory cell and APC function. A support protocol for collagenase digestion of splenocyte suspensions is described to increase the yield of dendritic cells. The second method involves generating large numbers of DCs from mouse bone marrow progenitor cells. - [Read Isolation of Dendritic Cells Protocol]

Isolation of Primary Epithelial Cells - http://www.lbl.gov/lifesciences/BissellLab/labprotocols/primarycellprep.htm

Category: Cell Biology and Cell Culture: Epithelial Cell Culture

Isolation of Primary Epithelial Cells. Digestion: In Collagenase Medium, Purification and epithelial cell culture methods. Bissell Lab. - [Read Isolation of Primary Epithelial Cells]

Molecular Cloning Lab â€“ Restriction Enzyme Digestion PDF - http://ender.bu.edu/~tgardner/be209/lab_manual/Lab%204.pdf

Category: Molecular Biology Protocols: Restriction Enzyme Digestion

Molecular Cloning Lab â€“ Restriction Enzyme Digestion. Molecular Cloning introduction, lab for students - restriction digest exercise with questions. BE209 Spring 2006. Prof. Timothy Gardner. Boston University, Department of Biomedical Engineering. - [Read Molecular Cloning Lab â€“ Restriction Enzyme Digestion PDF]

Articles (1-3 of 3)

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.

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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