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Preferential Delivery of the Sleeping Beauty transposon System to Livers of Mice by Hydrodynamic i - http://www.natureprotocols.com/2007/12/06/preferential_delivery_of_the_s.php

Category: Developmental Biology: Utilizing Model Organisms Protocols

Nonviral, DNA-mediated gene transfer is an alternative to viral delivery systems for expressing new genes in cells and tissues. The Sleeping Beauty (SB) transposon system combines the advantages of viruses and naked DNA molecules for gene therapy purposes; however, efficacious delivery of DNA molecules to animal tissues can still be problematic. Here we describe the hydrodynamic delivery procedure for the SB transposon system that allows efficient delivery to the liver in the mouse. - [Read Preferential Delivery of the Sleeping Beauty transposon System to Livers of Mice by Hydrodynamic i]

Preparation and LCM of Paraffin Embedded Tissue Sections Protocol - http://dir.nichd.nih.gov/lcm/LCMTAP.htm#II.%20Preparation%20and%20LCM%20of%20Paraffin%20Embedded%20Tissue%20Sections

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

Protocol for the preparation of LCM of Paraffin embedded tissue sections. Includes: Fixation; Processing, Embedding and Tissue Sectioning; Staining; Laser Transfer; DNA Extraction Protocol of LCM tissue and PCR for DNA analysis; RNA Extraction Protocol of LCM tissue and RT-PCR for mRNA analysis. - [Read Preparation and LCM of Paraffin Embedded Tissue Sections Protocol]

Preparing Siliconized Pipettes Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4355

Category: Mouse Protocols: Mouse Embryology Protocols

Protocol describes a method for preparing siliconized pipettes. Such pipettes minimize the loss of embryos or embryonic tissues during transfer. - [Read Preparing Siliconized Pipettes Protocol]

Production of Completely ES Cell-Derived Fetuses by Aggregation with Tetraploid Embryos - http://www.mshri.on.ca/nagy/Tetraploid/tetra.htm

Category: Stem Cell Protocols: Stem Cell Isolation Protocols

Protocol for the production of completely ES cell-derived fetuses by aggregation with tetraploid embryos. Includes: Recovery of 2-cell stage embryos; Production of tetraploid embryos; Preparation of aggregation plate; Removal of Zona Pellucida; ES cells/ tetraploid embryo "SANDWICH" aggregation; Transfer of embryos. - [Read Production of Completely ES Cell-Derived Fetuses by Aggregation with Tetraploid Embryos]

Protocol for Antiphosphotyrosine Western Blot Analysis - http://www.bio.com/protocolstools/protocol.jhtml?id=p697

Category: Signalling Signal Transduction Protocols

Detection of phosphorylated tyrosine residues can be performed using anti-P-TYR Ab and Western Analysis.Includes 2nd method,which uses phosphotyrosine in conjunction with anti-P-TYR Ab to "unlabel" potential proteins.By comparing Westerns developed with the 1st method(reveals phosphorylated protein) and the 2nd method(reveals non-specific labeling), a more accurate picture of those proteins phosphorylated on tyrosine can be seen. Includes: Protein Preparation, Electrophoresis and Transfer. - [Read Protocol for Antiphosphotyrosine Western Blot Analysis]

Removal of CCR5 Ligands and Induction of Pro-Resolving Lipid Mediators by Apoptotic Neutrophils - http://www.natureprotocols.com/2006/10/02/removal_of_ccr5_ligands_and_in.php

Category: Immunology

Removal of CCR5 ligands and induction of pro-resolving lipid mediators by apoptotic neutrophils during resolution. Application of lipid extraction from peritoneal exudates, in tandem with lipid mediator informatics can be used to determine the role of apoptotic neutrophils in the generation of resolution phase lipid mediators. This neutrophil transfer system allows the determination of the direct impact of apoptotic leukocytes in the resolution of inflammation. - [Read Removal of CCR5 Ligands and Induction of Pro-Resolving Lipid Mediators by Apoptotic Neutrophils]

Replication Timing by Density Transfer Protocol - http://fangman-brewer.genetics.washington.edu/density_transfer.html

Category: Yeast Protocols: Yeast Genetics Protocols: Yeast Replication Protocols

Protocol for replication timing by density transfer. - [Read Replication Timing by Density Transfer Protocol]

Root Transformation of Arabidopsis Protocol - http://www.cshprotocols.org/cgi/content/abstract/2006/30/pdb.prot4671

Category: Plant Biology Protocols: Arabidopsis Transformation Protocols

Arabidopsis can be stably transformed using Agrobacterium tumefaciens-mediated transfer of T-DNA. We describe the generation of transgenic plants via root transformation in tissue culture, which can be useful for transforming sterile mutants. - [Read Root Transformation of Arabidopsis Protocol]

Screening Bacterial Colonies by Hybridization: Intermediate Numbers Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3946

Category: Bacterial Protocols

Bacterial colonies growing on agar plates are transferred en masse to nitrocellulose filters. The spatial arrangement of colonies on the plates is preserved on the filters. After transfer, the filters are processed for hybridization to an appropriate radiolabeled probe while the original (master) plate is incubated for a few hours to allow the bacterial colonies to regrow in their original positions. - [Read Screening Bacterial Colonies by Hybridization: Intermediate Numbers Protocol]

Southern Blotting: Capillary Transfer of DNA to Membranes Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot4040

Category: Nucleic Acid Detection Protocols: Nucleic Acid Blotting Protocols

Protocol describes the first stages of Southern blotting: digestion of genomic DNA with one or more restriction enzymes, separation of the resulting fragments by electrophoresis through an agarose gel, and transfer of the denatured fragments to a membrane by downward capillary transfer. - [Read Southern Blotting: Capillary Transfer of DNA to Membranes Protocol]

Southern Blotting: Simultaneous Transfer of DNA from a Single Agarose Gel to Two Membranes Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot4043

Category: Nucleic Acid Detection Protocols: Nucleic Acid Blotting Protocols

Protocol for southern blotting: simultaneous transfer of DNA from a single agarose gel to two membranes. DNA can be simultaneously transferred from opposite sides of a single agarose gel to two membranes. Bidirectional transfer occurs rapidly at first, but soon slows down as the gel becomes dehydrated. Because the efficiency of transfer is low, the method works best when the target sequences are present in high concentration - [Read Southern Blotting: Simultaneous Transfer of DNA from a Single Agarose Gel to Two Membranes Protocol]

Staining Immunoblots for Total Protein Using India Ink Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/16/pdb.prot4270

Category: Western Blot Protocols: Immunoblot Stains for Total Protein Protocols

Protocol used to determine the position of molecular-weight markers and to ensure that efficient transfer of proteins to the blot has occurred, the total composition of the proteins can be determined by staining the membrane with Ponceau S (Staining Immunoblots for Total Protein Using Ponceau S) or India ink. - [Read Staining Immunoblots for Total Protein Using India Ink Protocol]

Staining Immunoblots for Total Protein Using Ponceau S Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/16/pdb.prot4269

Category: Western Blot Protocols: Immunoblot Stains for Total Protein Protocols

Protocol used to determine the position of molecular-weight markers and to ensure that efficient transfer of proteins to the blot has occurred, the total composition of the proteins can be determined by staining the membrane with Ponceau S . - [Read Staining Immunoblots for Total Protein Using Ponceau S Protocol]

TAIL PCR Protocol - http://preuss.bsd.uchicago.edu/protocols/tail.html

Category: DNA Protocols: PCR Polymerase Chain Reaction Protocols

TAIL PCR Protocol. TAIL is a series of reactions that are intended to map where a T-DNA (transfer DNA) has inserted within the genome. The main components of the 3 reactions are the AD (Arbitrary Degenerate) primers, border primers, and DNA from the T-DNA - [Read TAIL PCR Protocol]

TAIL PCR Protocol - http://preuss.bsd.uchicago.edu/protocols/tail.html

Category: PCR Protocols

Transfer and Fixation of Denatured RNA to Membranes Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot4060

Category: Nucleic Acid Electrophoresis Protocols: RNA Electrophoresis Protocols

Protocol describes the transfer of RNA from agarose gels to neutral or positively charged nylon membranes, using upward capillary flow of neutral or alkaline buffers. RNA becomes covalently fixed to positively charged nylon membranes during transfer in alkaline buffers. However, treatment by UV irradiation or heating is required to fix RNA to neutral membranes. - [Read Transfer and Fixation of Denatured RNA to Membranes Protocol]

Transfer of Bacteriophage DNA from Plaques to Filters Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3982

Category: Viral Manipulation Protocols: Phage Protocols

Protocol is used to identify and isolate recombinants containing DNA sequences of interest. - [Read Transfer of Bacteriophage DNA from Plaques to Filters Protocol]

Transfer of Eukaryote Suspension Cultures - http://homepages.gac.edu/~cellab/chpts/chpt12/ex12-6.html

Category: Cell Biology and Cell Culture

Transfer of Eukaryote Suspension Cultures. Cell Biology Laboratory Manual. Heidcamp. - [Read Transfer of Eukaryote Suspension Cultures]

Transfer of Eukaryote Suspension Cultures Protocol - http://homepages.gac.edu/~cellab/chpts/chpt12/ex12-6.html

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

Protocol for the transfer for eukaryote suspension cultures. - [Read Transfer of Eukaryote Suspension Cultures Protocol]

Transfer of Injected Blastocysts to Pseudo Pregnant Mice Protocol - http://www.bio.com/protocolstools/protocol.jhtml?id=p1118

Category: Mouse Protocols: Mouse Embryology Protocols

Blastocyst transfer is usually performed 24 hours after aggregation, when the morulae have become expanded blastocysts, and on the same day as injection. A little time is given between injection and transfer to allow the blastocysts to re-expand. Includes: The Mouse Recipient; The Transfer. - [Read Transfer of Injected Blastocysts to Pseudo Pregnant Mice Protocol]

Transfer of Phage DNA from Plaques to Nylon Membrane or Nitrocellulose Protocol - http://humgen.wustl.edu/hdk_lab_manual/lambda/lambda10.html

Category: Viral Culture and Isolation Protocols: Viral Culture Protocols

To transfer phage DNA to nylon membranes so that a phage library can be screened by hybridization. - [Read Transfer of Phage DNA from Plaques to Nylon Membrane or Nitrocellulose Protocol]

Transferring Bacteriophage P1 Clones between E. coli Hosts Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot4004

Category: Viral Manipulation Protocols: Phage Protocols

Protocol for transfer of P1 clones by electroporation. - [Read Transferring Bacteriophage P1 Clones between E. coli Hosts Protocol]

Troubleshooting Guide: Western Blot - http://www.rndsystems.com/tsg_detail_objectname_western_blot.aspx

Category: Protein Protocols: Western Blot Protocol: Troubleshooting Western Blots

Problem: Poor Transfer, Problem: Poor Transfer. R&D Systems. - [Read Troubleshooting Guide: Western Blot]

Uterine Transfer Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/18/pdb.prot4380

Category: Mouse Protocols: Mouse Reproductive System Surgery Protocols

Protocol describes a procedure for uterine transfer, which is used for chimera production. The method is based on extensive work which resulted in the first successful development and birth of in-vitro-cultured mouse embryos. It is best to practice this procedure first on a cadaver and then on an anesthetized 2.5-days post coitum (dpc) pseudopregnant mouse using blue Affigel beads rather than embryos. - [Read Uterine Transfer Protocol]

Vectors and Agrobacterium Hosts for Arabidopsis Transformation Protocol - http://www.cshprotocols.org/cgi/content/abstract/2006/30/pdb.ip29

Category: Plant Biology Protocols: Arabidopsis Transformation Protocols

A. tumefaciens is a soil-dwelling bacterium that transforms normal plant cells into tumor-forming cells by inserting a piece of bacterial DNA (the transfer, or "T," DNA) into the plant cell genome. The Ti plasmid also carries many of the transfer functions for mobilizing the T-DNA. This article provides a brief discussion of the principles of T-DNA transformation, including consideration of T-DNA vectors and their hosts. - [Read Vectors and Agrobacterium Hosts for Arabidopsis Transformation Protocol]

Articles (1-10 of 11)

Lambda Phage Protocol Large DNA Preparation

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.

Microarray Protocol Preparation of Fluorescent DNA Probes from Human mRNA

This Microarray Protocol Preparation of Fluorescent DNA Probes from Human mRNA protocol describes the production of probes labeled with the fluorescent dyes, Cy3 and Cy5, following the synthesis of cDNA from human mRNA and the hybridization of the probes to DNA microarrays.

Probe Preparation for 22 x 40 mm DNA Microarrays

Probe Preparation for 22 x 40 mm DNA Microarrays Protocol.

Acid Guanidinium Thiocyanate RNA Isolation Phenol Chloroform Extraction

A single step RNA isolation protocol using Phenol Chloroform Extraction and Acid Guanidinium Thiocyanate. This RNA isolation method uses the fact that guanidinium thiocyanate can simultaneously lyse the cells and inactive cellular RNAses during the initial RNA isolation step allow a single step in the method.

In Vitro Translated Xenopus Mos Kinase Assay Protocol

In Vitro Translated Xenopus Mos Kinase Assay Protocol. In response to progesterone, immature Xenopus oocytes mature to eggs that can be fertilized. The Mos protein kinase is essential for oocyte maturation, most likely due to its ability to activate the MAP kinase cascade. This MAP kinase cascade eventually leads to the activation of Cdc2/cyclin B and entry into M phase. In this protocol, tagged Mos kinase is translated in vitro, immunopurified, and used in a kinase assay.

Antibodies Phage Expression Protocol Using 96-well Microtiter Plates

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

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.