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Sequence Specific Retention Calculator - http://hs2.proteome.ca/SSRCalc/SSRCalc.html

Sequence Specific Retention Calculator. A sequence-specific correction factors for prediction of peptide retention in RP-HPLC: application to protein identification by off-line HPLC-MALDI-MS. Oleg Krokhin - [Read Sequence Specific Retention Calculator]

Transfection of Bone Marrow-Derived Mast Cells for Transcription Factor Luciferase Reporter Assays - http://www.natureprotocols.com/2006/09/12/transfection_of_bone_marrowder_1.php

Category: Reporter Gene Assays: Luciferase Assay Protocols

Transfection of primary leukocytes has traditionally been a challenging but much desired protocol. It allows not only the analysis of cells in a more natural state to a cell line system, it enables the direct comparison of, for e.g. transcriptional activity using luciferase reporters, in immune cells taken from genetically-altered mice. In addition, importantly it allows for "rescue experiments" in knockout cells & the ability to over-express or reconstitute wild-type and/or mutated constructs. - [Read Transfection of Bone Marrow-Derived Mast Cells for Transcription Factor Luciferase Reporter Assays]

Transient Transfection Into 293T Cells Protocol - http://www.cbrinstitute.org/labs/springer/protocols/jun_293T.html

Category: Transfection Protocols: Transient Transfection Protocols

Transient transfection into 293T cells is a convenient way to overexpress and obtain both cellular and extracellular (secreted or membrane) proteins. 293 is a human renal epithelial cell line which is transformed by adenovirus E1A gene product. 293T is a derivative which also express SV40 large T antigen, allowing episomal replication of plasmids containing the SV40 origin and early promoter region. They (both) have the unusual property of being highly transfectable. - [Read Transient Transfection Into 293T Cells Protocol]

Transient Transfection Protocol: Viral Production - http://genetics.med.harvard.edu/~cepko/protocol/mike/V2.html

Category: Transfection Protocols: Transient Transfection Protocols

Transient transfection protocol for the ecotropic/amphotropic producer line including titering and concentration of viral stocks. - [Read Transient Transfection Protocol: Viral Production]

Transient Transfection: Viral Production - http://genetics.med.harvard.edu/~cepko/protocol/mike/V1.html

Category: Transfection Protocols: Transient Transfection Protocols

This transient transfection protocol for the ecotropic/amphotropic producer line includes instructions for titering and concentration of viral stocks. This procedure gives consistently higher transfection efficiency. - [Read Transient Transfection: Viral Production]

Trypsinization of Cells Grown in Monolayer Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4349

Category: Cell Culture Protocols: Trypsinization Protocols

Protocol describes the trypsinization of cells in monolayer culture to facilitate subculture or harvesting. To avoid cross-contamination of cells, it is important for each cell line to be subcultured independently. No more than one cell line should be in the tissue culture hood at any one time. - [Read Trypsinization of Cells Grown in Monolayer Protocol]

X-Ray Mutagenesis of Drosophila Protocol - http://www.bioprotocol.com/protocolstools/protocol.jhtml%3Bjsessionid%3DGYZOPNVU3IQQTR3FQLMSFEWHUWBNQIV0?id=p32

Category: Drosophila Protocols: Drosophila Genetics Protocols

Protocol describes the general procedure for creating mutations in the DNA of Drosophila by exposure to X-rays. Irradiation of cells with X-rays creates double strand breaks (DSBs) in DNA. Mutations introduced in the DNA of germ line cells (sperm) are propagated by mating the exposed males to virgin females. The progeny of this cross can be mated to each other so that a percentage of the subsequent offspring will have two copies of the same mutant allele. - [Read X-Ray Mutagenesis of Drosophila Protocol]

Articles (1-10 of 16)

Single Stranded Plasmid DNA Isolation Protocol

A Single Stranded Plasmid DNA Isolation Protocol describing the production and isolation of single-stranded DNA (ssDNA) using bacteriophagemid-containing bacteria and helper phage. Infection of the host cells with helper phage allows for packaging of ssDNA into bacteriophage. The ssDNA can then be isolated from phage particles.

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.

Drosophila Cell Transfection Protocol

A simple Drosophila tissue culture cell transfection method.

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.

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.

Acid Washing Microinjection Pipettes Protocol

Acid Washing of Glass microinjection pipettes protocol.

Paraffin Embedding Protocol

Paraffin Embedding Protocol for molecular profiling. This Paraffin Embedding Protocol describes the processing of the tissues into sections following ethanol fixation. Molecular profiling (MP) is a technique that is used to visualize the global patterns of RNA expression or protein expression in various cell types and disease processes.

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.