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3' Rapid Amplification of cDNA Ends (RACE) Protocol - http://www.nottingham.ac.uk/~mbzspd/methods/3RACE_PCR.html

Category: RNA Protocols: 3' RACE Rapid Amplification of cDNA Ends Protocols

PCR cycle steps, protocol using SuperScript II reverse transcriptase (Life Technologies) and Taq polymerase, Gene specific primer, T17 Adapter primer and an Adapter primer. Dr.Dawson, Nottingham. - [Read 3' Rapid Amplification of cDNA Ends (RACE) Protocol]

A Rapid Method for Generating Gene Deletions in Aspergillus fumigatus Protocol - http://www.aspergillus.org.uk/indexhome.htm?secure/laboratory_protocols/./genedel.html~main

Category: Fungi Protocols: Aspergillus Protocols

The technique makes use of an Escherichia coli strain expressing the redΑßΓ operon under the control of an inducible promoter. This enables the strain to carry out homologous recombination with only 50-60 bp of homologous sequence. The procedure does not require any DNA ligation and is very rapid. It allows a single gene or region on a cosmid to be replaced by a bi-functional selectable marker (having both an E. coli and an A. fumigatus marker). - [Read A Rapid Method for Generating Gene Deletions in Aspergillus fumigatus Protocol]

A Rapid Method for Isolation of Total Nucleic Acids from Aspergillus nidulans Protocol - http://www.fgsc.net/fgn/chow.html

Category: Fungi Protocols: Aspergillus Protocols

Protocol for the rapid method for isolation of total nucleic acids from Aspergillus nidulans. - [Read A Rapid Method for Isolation of Total Nucleic Acids from Aspergillus nidulans Protocol]

A Simplified System for Rapid Generation of Recombinant Adenoviruses - http://www.coloncancer.org/adeasy/protocol.htm

Category: Viral Culture and Isolation Protocols: Viral Isolation Protocols

A simplified system for rapid generation of recombinant adenoviruses. Includes: Generation of Recombinants in Bacterial Cells; Viral Production in 293 or 911 Cells; Preparation of High Titer Viral Stocks. - [Read A Simplified System for Rapid Generation of Recombinant Adenoviruses]

A Strategy for the Rapid Identification of - http://www.people.virginia.edu/~wrp/papers/mcp_wrp02b.pdf

Category: Protein Protocols: Protein Phosphorylation Protocols

A Strategy for the Rapid Identification of Phosphorylation Sites in the Phosphoproteome. MacDonald et al., Molecular & Cellular Proteomics. 2002. - [Read A Strategy for the Rapid Identification of]

AFLP genotyping and fingerprinting Review PDF - http://taxonomy.zoology.gla.ac.uk/~rdmp1c/teaching/L4/Evolution/Session5/Mueller.pdf

Category: PCR Protocols: AFLP PCR

Ulrich G. Mueller and L. LaReesa Wolfenbarger. Amplified fragment length polymorphisms (AFLPs) are polymerase chain reaction (PCR)-based markers for the rapid screening of genetic diversity. AFLP. TREE October 1999 - [Read AFLP genotyping and fingerprinting Review PDF]

Allium Test - http://embryo.ib.amwaw.edu.pl/invittox/prot/8.htm

Category: Pharmacology and Toxicology Protocols: Cytotoxicity Protocols

The Allium test provides a rapid screening procedure for chemicals, pollutants contaminants, etc. which may represent environmental hazards. Root growth inhibition and adverse effects upon chromosomes provide an indication of likely toxicity. - [Read Allium Test]

Allium Test - http://embryo.ib.amwaw.edu.pl/invittox/prot/8.htm

Category: Plant Biology Protocols

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]

Bisulfite-PCR for Restriction Analysis and/or Sequencing Protocol - http://www.mdanderson.org/departments/methylation/display.cfm?id=8E0AAAAC-FA42-486C-A93F60CD6E1C7E51&method=displayFull&pn=A3F15D82-C9B4-41CD-BA4BE767E50A73D2

Category: PCR Protocols: Bisulfite Conversion Based PCR Protocols

Protocol for bisulfite-PCR for restriction analysis and/or sequencing. Bisulfite-PCR followed by restriction is a rapid and semi-quantitative method of analyzing DNA methylation. The PCR products are also suitable for either direct sequencing or cloning and sequencing. The most important step here is primer selection. - [Read Bisulfite-PCR for Restriction Analysis and/or Sequencing Protocol]

DNA Isolation From BAC & PAC Clones Protocol - http://bacpac.chori.org/bacpacmini.htm

Category: BACs Bacterial Artificial Chromosome Protocols

Rapid alkaline lysis miniprep method for isolating DNA from large PAC clones. It is a modification of a standard Qiagen-Tip method that uses no organic extractions or columns. The method works very well for doing analytical restriction digests of PAC clones and can be scaled up if necessary. - [Read DNA Isolation From BAC & PAC Clones Protocol]

DNA Isolation From BAC & PAC Clones Protocol - http://bacpac.chori.org/bacpacmini.htm

Category: Nucleic Acid Purification Protocols: DNA Isolation Protocols

This is a rapid alkaline lysis miniprep method for isolating DNA from large PAC clones. It is a modification of a standard Qiagen-Tip method that uses no organic extractions or columns. The method works very well for doing analytical restriction digests of PAC clones and can be scaled up if necessary. - [Read DNA Isolation From BAC & PAC Clones Protocol]

Enzyme Immunoaffinity Chromatographyâ€”A Rapid Semi-quantitative Method - http://www.rsc.org/delivery/_ArticleLinking/DisplayArticleForFree.cfm?doi=a607578i

Category: Chromatography Protocols: Immunoaffinity Chromatography Protocols

Enzyme Immunoaffinity Chromatographyâ€”A Rapid Semi-quantitative Method. - [Read Enzyme Immunoaffinity Chromatographyâ€”A Rapid Semi-quantitative Method]

Epitope Mapping Using Synthetic Biotin-Labeled Peptides Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/18/pdb.prot4278

Category: Antibody Protocols: Epitope Mapping Antibody Protocols

A set of overlapping synthetic peptides is synthesized, each corresponding to a small segment of the linear sequence of a protein antigen and arrayed on a solid phase. The panel of solid-phase peptides is then probed with a test antibody, and bound antibody is detected using an enzyme-labeled secondary antibody. This method is very rapid and can be extraordinarily successful. - [Read Epitope Mapping Using Synthetic Biotin-Labeled Peptides Protocol]

Extraction of Bacteriophage lambda DNA from Large-scale Cultures Using Formamide Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3973

Category: Viral Manipulation Protocols: Phage Lamda Manipulation Protocols

Formamide can be used instead of proteinase K to dissociate bacteriophage {lambda} coat proteins from the viral {lambda}. The procedure is rapid and works best with large-scale preparations of bacteriophage {lambda}. - [Read Extraction of Bacteriophage lambda DNA from Large-scale Cultures Using Formamide Protocol]

Extraction of Calf Brain Lipids Protocol - http://www.bio.com/protocolstools/protocol.jhtml?id=p1281

Category: Lipid Protocols

Protocol for the extraction of calf brain lipids. Protocol describes a rapid method to isolate lipids from bovine brain tissue using an organic solvent mixture of Chloroform and Methanol. - [Read Extraction of Calf Brain Lipids Protocol]

Fluorescent In Situ Transcription in Cells and Tissues Protocol - https://catalog.invitrogen.com/index.cfm?fuseaction=iProtocol.unitSectionTree&treeNodeID=9E6639A7FEDD7A49BBC3758516F99557&objectid=6677C9E8B913BFB72E9F654025F3C39A

Category: Cytogenetics Protocols: Fluorescent In Situ Hybridization FISH Protocol

Method assesses cellular mRNA transcripts in tissue sections and cell cultures using unique short anti-sense primers directed against sequences in particular protein(s). The unlabeled synthetic cDNA oligonucleotide primers are extended complementary to a sense mRNA transcript using reverse transcriptase and labeled through incorporation of a fluorescent-labeled dUTP nucleotide base. This procedure provides rapid detection of low abundance mRNA messages that can be related to other cellular.... - [Read Fluorescent In Situ Transcription in Cells and Tissues Protocol]

G1 Synchronization of CHO Cells by Isoleucine Deprivation Protocol - http://www.cshprotocols.org/cgi/content/abstract/2006/23/pdb.prot4486

Category: Cell Biology and Cell Culture: Cell Cycle Protocols: G Phase Protocols

This protocol provides a method for the synchronization of a monolayer culture of CHO cells in G1 using isoleucine deprivation. Since CHO cells can also be adapted to grow in suspension culture, this procedure can be used to obtain larger quantities of cells. When isoleucine is replaced, the cells resume growth and begin to enter S phase ~4 hours later. This method arrests almost 100% of the CHO cells in G1, and upon reversal, leads to rapid recovery of cell growth and very high cell viability. - [Read G1 Synchronization of CHO Cells by Isoleucine Deprivation Protocol]

G1 Synchronization of CHO Cells by Isoleucine Deprivation Protocol - http://www.cshprotocols.org/cgi/content/abstract/2006/23/pdb.prot4486

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

Genotyping Embryonic Stem (ES) Cell Colonies Prior to Picking Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/21/pdb.prot4415

Category: Genetics and Genomics: Genotyping Protocols: PCR Genotyping

This protocol describes a rapid PCR-based method for identifying targeted ES cell colonies prior to picking. It is based on DNA analysis of a small part of colonies pooled directly from selection plates. Only positive colonies are expanded. - [Read Genotyping Embryonic Stem (ES) Cell Colonies Prior to Picking Protocol]

Identifying Transformed Arabidopsis thaliana Seedlings Following Floral Dip Transformation Protocol - http://www.plantmethods.com/content/2/1/19

Category: Plant Biology Protocols: Arabidopsis Transformation Protocols

Protocol for a rapid and robust method of identifying transformed Arabidopsis thaliana seedlings following floral dip transformation. - [Read Identifying Transformed Arabidopsis thaliana Seedlings Following Floral Dip Transformation Protocol]

Immuno-Laser Capture Microdissection Protocol - http://cgap-mf.nih.gov/Protocols/Slides/SlideProtocols/ImmunoLCM.html

Category: Primary Cell Isolation and Culture Protocols: Laser Capture Microdissection Protocols

After fixation, frozen sections are immunostained under RNase-free conditions using a rapid three-step streptavidin-biotin technique followed by dehydration. The immunostained sections are ready for LCM. Includes: Development of Immuno-LCM. - [Read Immuno-Laser Capture Microdissection Protocol]

Introduction to Live-Cell Imaging Techniques - http://microscopy.fsu.edu/primer/techniques/livecellimaging/index.html

Category: Microscopy Protocols: Live-Cell Imaging Protocols

Live-cell imaging techniques provide critical insight into the fundamental nature of cellular & tissue function, especially due to the rapid advances that are currently being witnessed in fluorescent protein & synthetic fluorophore technology. Because of these advances, live-cell imaging has become a requisite analytical tool in most cell biology labs. Includes: Maintaining Live Cells on the Microscope Stage; Live-Cell Imaging Culture Chambers; Optical System and Detector Requirements etc. - [Read Introduction to Live-Cell Imaging Techniques]

Isolation of High-molecular-weight DNA from Mammalian Cells Using Proteinase K and Phenol Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot4036

Category: Nucleic Acid Purification Protocols: DNA Isolation Protocols: DNA Isolation from Mammalian Cells Protocols

Method of choice when large amounts of mammalian DNA are required, for example, for Southern blotting (Rapid Isolation of Mammalian DNA, Rapid Isolation of Yeast DNA, Southern Blotting: Capillary Transfer of DNA to Membranes) or for construction of genomic libraries in bacteriophage {lambda} vectors. Approximately 200 µg of mammalian DNA, 100-150 kb in length, is obtained from 5 x 107 cultured aneuploid mammalian cells (e.g., HeLa cells). - [Read Isolation of High-molecular-weight DNA from Mammalian Cells Using Proteinase K and Phenol Protocol]

Kit for Rapid Enrichment of SARS Virus - http://www.gentaur.com/sars_isolation.htm

Category: Viral Culture and Isolation Protocols: Viral Isolation Protocols

During incubation period and the course of disease, SARS virus replicates and releases from cells. The level of virus varies greatly from sample to sample. At early stage of incubation period or during the late convalescent phase, the concentration of virus is very low in all kinds of samples. Virus is also detected at extremely low concentrations in plasma during... - [Read Kit for Rapid Enrichment of SARS Virus]

Articles (1-3 of 3)

Tubulin Polymerization Protocol using GTP and GMPCPP

Tubulin is polymerized into microtubules by incubating tubulin at 37°C with GTP. A nucleation seed is added when the purpose is to assay microtubule elongation. Tubulin can also be polymerized for the purposes of recycling the tubulin or labeling the microtubules with fluorescently labeled tubulin. Based on the protocol by Timothy Mitchison of Harvard University.

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.

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