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Restriction enzyme digestion of DNA: basic method - http://www.methodbook.net/dna/restrdig.html

Category: Molecular Biology Protocols: Restriction Enzyme Digestion

How much DNA to digest? Master mix for restriction digest. Double digests. Star activity. Matt Lewis, Department of Pathology, Univ. Liverpool. - [Read Restriction enzyme digestion of DNA: basic method]

Analysis of DNA by Restriction Enzyme Cleavage and Agarose Gel Electrophoresis - http://www.ambl.msl.ubc.ca/exp2.html

Category: Molecular Biology Protocols: Restriction Enzyme Digestion

Very detailed class protocol for Analysis of DNA by Restriction Enzyme Cleavage and Agarose Gel Electrophoresis. David Ng, UBC. - [Read Analysis of DNA by Restriction Enzyme Cleavage and Agarose Gel Electrophoresis]

Enzyme Purification by Acetone Precipitation - http://www.glue.umd.edu/~nsw/ench485/lab6b.htm

Category: Protein Protocols: Protein Precipitation Procols

Acetone Precipitation Protocol and Method. Nam Sun Wang. Univ. Maryland. - [Read Enzyme Purification by Acetone Precipitation]

Enzyme Purification by Salt Ammonium Sulfate Precipitation - http://www.ece.umd.edu/~nsw/ench485/lab6a.htm

Category: Protein Protocols: Protein Precipitation Procols

ENZYME PURIFICATION BY SALT (AMMONIUM SULFATE) PRECIPITATION. Prepared by Nam Sun Wang, Department of Chemical & Biomolecular Engineering. University of Maryland. - [Read Enzyme Purification by Salt Ammonium Sulfate Precipitation]

ELISA (Enzyme-Linked ImmunoadSorbent Assay) - http://www.nottingham.ac.uk/~mbzspd/methods/ELISA.html

Category: ELISA Protocols

ELISA (Enzyme-Linked ImmunoadSorbent Assay) Method and technique. Jill Fergusson, Department of Biochemistry, University of Nottingham, - [Read ELISA (Enzyme-Linked ImmunoadSorbent Assay)]

Restriction Digestion Methods - http://hg.wustl.edu/hdk_lab_manual/rdm/rdmcontents.html

Category: Molecular Biology Protocols: Restriction Enzyme Digestion

Restriction Digestion Methods. Donis-Kellor lab manual. Restriction Enzyme Digests, Restriction Enzyme Buffers, Restriction Digestion of Plasmid, Cosmid, and Phage DNAs. - [Read Restriction Digestion Methods]

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]

Restriction Enzyme Digestion of DNA - http://www.facstaff.bucknell.edu/pizzorno/327-lab4.html

Category: Molecular Biology Protocols: Restriction Enzyme Digestion

Digesting DNA with Restriction Endonucleases introduction. Analytical and Preparative reaction mixes. All about restriction enzymes. Pizzorno, BIOL 327. - [Read Restriction Enzyme Digestion of DNA]

Cytokine ELISA Protocol - http://www.bdbiosciences.com/pharmingen/protocols/Cytokine_ELISA.shtml

Category: ELISA Protocols: Sandwich ELISA Assay Protocols

Cytokine sandwich ELISA are sensitive enzyme immunoassays that can specifically detect and quantitate the concentration of soluble cytokine and chemokine proteins. BD Biosciences - [Read Cytokine ELISA Protocol]

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]

Assay for ß-galactosidase in Extracts of Mammalian Cells - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3952

Category: Reporter Gene Assays: B-Galactosidase Assay Protocols

The assay for ß-galactosidase relies on the ability of the enzyme to catalyze the hydrolysis of ONPG (o-nitrophenyl-ß-D- galactopyranoside) to free o-nitrophenol, which absorbs light at 420 nm. In this protocol, extracts of cells transfected with a ß-galactosidase reporter plasmid are incubated with ONPG. - [Read Assay for ß-galactosidase in Extracts of Mammalian Cells]

Apoptosis Detection Using TUNEL Enzymatic - http://www.ihcworld.com/_protocols/apoptosis/tunel_enzyme.htm

Category: Cell Biology and Cell Culture: Apoptosis Protocols: TUNEL assay apoptosis

Apoptosis Detection Using Terminal Transferase and Biotin-16-dUTP (TUNEL -Enzyme Method). IHC World - [Read Apoptosis Detection Using TUNEL Enzymatic]

Restriction Enzyme Troubleshooting Guide - http://www.fermentas.com/techinfo/re/troubleshoot.htm

Category: Molecular Biology Protocols: Restriction Enzyme Digestion

Restriction Enzyme Troubleshooting Guide. Fermentas. Problems: Undigested or incompletely digested DNA, additional bands atypical, Diffused DNA zones, Low fragment ligation efficiency. - [Read Restriction Enzyme Troubleshooting Guide]

Cloning Enzymes a Guide Promega - http://www.promega.com/guides/cloning_guide/default.htm

Category: Molecular Biology Protocols: Restriction Enzyme Digestion

Cloning Enzymes a Guide Promega. An Enzyme Resource Guide series, highlights those enzymes important in nucleic acid cloning procedures. Promega - [Read Cloning Enzymes a Guide Promega]

Fan Lab Bisulfite Treatment of Genomic DNA - http://www.genetics.ucla.edu/labs/fan/Protocols_Bisulfite.htm

Category: Epigenetics and Methylation Protocols: Bisulfite Treatment Methylation Assay Protocol

A detailed protocol for bisulfite treatment of DNA. DNA is first digested with a restriction enzyme. DNA is denauted at 97C for 5min. Bisulfite protocol is included. Also DNA purification using the Promega Wizard DNA Cleanup kit in indicated. Fan Lab. UCL - [Read Fan Lab Bisulfite Treatment of Genomic DNA]

Blocking Solutions Protocols - http://icg.cpmc.columbia.edu/cattoretti/Protocol/main_page_reagents.html#Blocking%20solutions

Category: Immunohistochemistry Protocols: Blocking Protocols

Protocols for blocking solutions. Includes: Endogenous peroxidase blocking solution (H2O2-Azide); Non-specific antibody background blocking solution: swine serum; Non-specific antibody background blocking solution: MILK; Enzymatic antigen retrieval solution; Unconjugated, biotin- and fluorochrome-conjugated antibody (primary) solutions; Unconjugated, biotin- and fluorochrome-conjugated antibody (secondary) solutions; Enzyme-conjugated secondary antibody solutions.; etc.. - [Read Blocking Solutions Protocols]

Blunt-end Cloning of PCR Products Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3830

Category: PCR Protocols: PCR Cloning Protocols

Protocol for blunt-end cloning of PCR products. Incubation of a blunt-end ligation reaction in the presence of an excess amount of an appropriate restriction enzyme can dramatically increase the yield of recombinant plasmids. The role of the restriction enzyme is to cleave circular and linear concatemers at restriction sites that are re-formed when linear, blunt-ended plasmid molecules ligate to themselves. I - [Read Blunt-end Cloning of PCR Products Protocol]

A specific endpoint assay for ubiquitin. - http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=304457

Category: Protein Protocols: Protein Ubiquitination Protocols

A specific endpoint assay for ubiquitin. Rose et al., Proc Natl Acad Sci U S A. 1987. Simple endpoint assays for free ubiquitin (Ub) and for the Ub-activating enzyme are described. . - [Read A specific endpoint assay for ubiquitin.]

Photosynthesis and Respiration - Introduction - http://homepages.gac.edu/~cellab/chpts/chpt8/intro8.html

Category: Plant Biology Protocols: Photosynthesis and Respiration Protocols

The physiological reactions of mitochondria and chloroplasts can be reduced to a series of electron transfers, catalyzed by specific enzymes found within the organelles. Thus, we can study the component processes of photosynthesis and respiration by isolating the organelles and measuring specific enzyme activity associated with that organelle. - [Read Photosynthesis and Respiration - Introduction]

ELISA for Antibody Isotype Determination Protocol - http://www.natureprotocols.com/2007/01/22/immunizations_and_enzymelinked.php

Category: ELISA Protocols

Protocol for immunizations and enzyme-linked immunosorbent assay (ELISA) for antibody isotype determination. - [Read ELISA for Antibody Isotype Determination Protocol]

Rebase The Restriction Enzyme Database - http://rebase.neb.com/rebase/rebase.html

Category: Molecular Biology Protocols: Restriction Enzyme Digestion

REBASE - The Restriction Enzyme Database - [Read Rebase The Restriction Enzyme Database]

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]

Shrimp Alkaline Phosphatase Protocol - http://www.usbweb.com/brief_proto/70092b.pdf

Category: DNA Protocols: SAP Protocols

Shrimp Alkaline Phosphatase Protocol. USB. Protocol for Dephosphorylation of 5'-ends of DNA in Restriction Enzyme Reaction. Protocol for Dephosphorylation of 5'-ends of DNA. - [Read Shrimp Alkaline Phosphatase Protocol]

ELISA for detection of platelet-associated Ig (PAIg) - http://www.icp.ucl.ac.be/mexp/vipg_plelisa.html

Category: ELISA Protocols

Qualitative ELISA (Enzyme-Linked Immunosorbent Assay) used for screening detection of anti-platelet antibodies or for detection of platelet-associated Ig (PAIg). Andrei Musaji Viral immunity and pathogenesis group, Universite Catholique de Louvain. - [Read ELISA for detection of platelet-associated Ig (PAIg)]

Apoptosis Detection: TUNEL Protocol - http://biochem.dental.upenn.edu/~sdmfacs/apopproc.htm#brduapop

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols: Apoptosis Cytometry Protocols

Protocol for apoptosis detection using TUNEL protocol. This technique uses the enzyme terminal deoxynucleotidyl transferase (TdT) to label cells that have oligonucleosomal nicks/strand breaks in their DNA. - [Read Apoptosis Detection: TUNEL Protocol]

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Populational Analysis of Genomic DNA Protocol

A protocol on the populational Analysis of genomic DNA.

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.

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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