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Harvesting the mRNA for Exon Trapping and Amplification - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3734

Category: DNA Protocols: Genomic DNA Library Protocols

Standard techniques are used here to isolate cytoplasmic RNA from COS cells transfected with a library of recombinant plasmids containing the genomic DNA of interest. - [Read Harvesting the mRNA for Exon Trapping and Amplification]

Histotechniques Fixation - http://library.med.utah.edu/WebPath/HISTHTML/HISTOTCH/HISTOTCH.html

Category: Immunohistochemistry Protocols: Fixation Protocols

Information on histotechniques. Includes: Fixation - types of fixatives; factors affecting fixation; General usage of fixatives; Tissue Processing; Sectioning; Frozen Sections; Staining; H and E staining; Coverslipping; Decalcification; Artefacts in Histologic Sections; Problems in Tissue Processing. - [Read Histotechniques Fixation]

Holzer's Stain Protocol - http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/HOLZER.PDF

Category: Neuroscience Protocols: Neuronal Stains Protocols

An appropriate term for glial fibers is 'nerve glue', because they provide the internal support of the central nervous system. There are four types of glial cells: astrocytes, oligosendroglia,microglia, and ependymal cells. The glia fibers are stained with crystal violet which are resistant to the aniline-chloroform differentiating solution. - [Read Holzer's Stain Protocol]

Immunoscreening of Lambda-GT11 Bacteriophage Libraries with Antibody Probes - http://omrf.ouhsc.edu/~frank/GT11.html

Category: RNA Protocols: cDNA Libraries Library

Adsroption of Anti-E. coli Antibodies, cDNA Library Screening, Frank Lab. Oklahoma Medical Research Foundation - [Read Immunoscreening of Lambda-GT11 Bacteriophage Libraries with Antibody Probes]

Isolation of Arabidopsis cDNAs by Complementation in Yeast - http://www.arabidopsis.org/cshl-course/12-yeast.html

Category: Plant Biology Protocols: Arabidopsis Nucleic Acid Protocols

Includes: Isolation of Arabidopsis LEU2 cDNAs by complementation of the yeast leu2 mutation; Recovery of plasmid DNA from yeast cells; Analysis of Arabidopsis cDNAs that complement a yeast leu2 mutation; Preparation of library DNA from the Lacroute cDNA library; Preparation of Yeast Media. - [Read Isolation of Arabidopsis cDNAs by Complementation in Yeast]

Isolation of Plant Transcription Factors Using a Modified Yeast One-Hybrid System - http://www.plantmethods.com/content/2/1/3

Category: Plant Biology Protocols: Plant Genetics Protocols

The preparation of expressional cDNA libraries for use in the yeast two-hybrid system is quick and efficient when using the dedicated Clontech™ product, the MATCHMAKER Library Construction and Screening Kit 3. This kit employs SMART technology for the amplification of full-length cDNAs, in combination with cloning using homologous recombination. - [Read Isolation of Plant Transcription Factors Using a Modified Yeast One-Hybrid System]

Library cDNA Synthesis - http://www.nottingham.ac.uk/~mbzspd/methods/Library_cDNA_Synthes.html

Category: RNA Protocols: cDNA Libraries Library

Library cDNA Synthesis. 1rst and 2nd strand cDNA synthesis. Dr. Simon Dawson - [Read Library cDNA Synthesis]

Ligation of Bacteriophage lambda Arms to Fragments of Foreign Genomic DNA Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3980

Category: Viral Manipulation Protocols: Phage Lamda Manipulation Protocols

Pilot ligations and packaging reactions are used to establish the amounts of fragmented genomic DNA and bacteriophage {lambda} arms that yield the maximum number of recombinants. Additional ligation and packaging reactions may then be set up to yield a comprehensive library of genomic DNA. - [Read Ligation of Bacteriophage lambda Arms to Fragments of Foreign Genomic DNA Protocol]

Ligation of cDNA to Bacteriophage lambda Arms for the Construction of cDNA Libraries - http://molecularstation.com/protocol-links/admin/dir_links_edit.php?action=N

Category: DNA Protocols: cDNA Library Protocols

The final stage of cDNA library construction involves optimizing ligation of the size-fractionated cDNA to bacteriophage {lambda} arms, followed by packaging and plating of the recombinant bacteriophages. - [Read Ligation of cDNA to Bacteriophage lambda Arms for the Construction of cDNA Libraries]

Method: Transfer of DNA from Bacterial Colonies to Nylon Membrane or Nitrocellulose - http://humgen.wustl.edu/hdk_lab_manual/plasmid/plsmid04.html

Category: RNA Protocols: cDNA Libraries Library

Purpose: To transfer DNA from bacterial colonies to nylon (or nitrocellulose) membrane so that a library can be screened by hybridization. Jim Howe Donis Keller Lab - [Read Method: Transfer of DNA from Bacterial Colonies to Nylon Membrane or Nitrocellulose]

Mixed Oligonucleotide-primed Amplification of cDNA MOPAC Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3484

Category: PCR Protocols: cDNA Synthesis Protocols

In MOPAC, the amino-terminal and carboxy-terminal sequences of a peptide are used to design two redundant families of oligonucleotides encoding the aminoand carboxy-terminal sequences of the peptide. The primers are used either to amplify a segment of cDNA prepared by RT-PCR from a tissue known to express the protein or to amplify a segment of DNA from an established genomic or cDNA library. - [Read Mixed Oligonucleotide-primed Amplification of cDNA MOPAC Protocol]

Modified Thioflavin for Senile Plaques Amyloid Protocol - http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/MODTHIOF.PDF

Category: Neuroscience Protocols: Neuronal Stains Protocols: Stains for Senile Plaques Protocols

Protocol for modified thioflavin for senile plaques amyloid. - [Read Modified Thioflavin for Senile Plaques Amyloid Protocol]

Neuroanatomy Tutorial - http://library.med.utah.edu/WebPath/HISTHTML/NEURANAT/NEURANCA.html

Category: Neuroscience Protocols

This tutorial has images in which the structures are labelled. You are to identify the structures by clicking on the name of the structure. - [Read Neuroanatomy Tutorial]

Partial Digestion of Eukaryotic DNA for Use in Genomic Libraries: Pilot Reactions Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3978

Category: Nucleic Acid Purification Protocols: DNA Isolation Protocols: Genomic DNA Isolation Protocols

Protocol is used to establish conditions for restriction enzyme digestion of eukaryotic genomic DNA that will generate fragments of a size appropriate for construction of genomic libraries. To construct a genomic library, the average length of the starting genomic DNA should be at least eight times the capacity of the vector. - [Read Partial Digestion of Eukaryotic DNA for Use in Genomic Libraries: Pilot Reactions Protocol]

PCR based Lambda Î»DNA library screening method - http://www.methods.info/Methods/Library_screening/library_pcr.pdf

Category: RNA Protocols: cDNA Libraries Library

"useful screen phage library non-radioactive, and not need too many clones. Up to 5 clones can be obtained usually within one week." Methods.info - [Read PCR based Lambda Î»DNA library screening method]

PCR-Based Screening of DNA Libraries Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/14/pdb.prot4129

Category: DNA Protocols: cDNA Library Protocols: cDNA Library Construction Protocols

PCR-Based Screening of DNA Libraries Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/14/pdb.prot4129

Category: PCR Protocols

PCR can be used to identify rare DNA sequences in DNA libraries by increasing the abundance of a particular sequence. This is accomplished by subdividing the original library into pools of decreased complexity and screening each pool or group of pools for a given DNA sequence. A pool that contains the desired clone is subsequently subdivided into smaller pools, each of which is screened using the same PCR protocol that was used for the primary screen. - [Read PCR-Based Screening of DNA Libraries Protocol]

Performing a Hunt by Interaction Mating Protocol - http://proteome.wayne.edu/Matinghunt.html

Category: Yeast Protocols: Yeast Genetics Protocols

When more than one bait will be used to screen a single library, significant time and resources can be saved by performing the interactor hunt by interaction mating. In this protocol one strain is transformed with library DNA and the transformants are collected and frozen in aliquots. - [Read Performing a Hunt by Interaction Mating Protocol]

Ph.D.â„¢-12 Phage Display Peptide Library Kit - http://www.neb.com/nebecomm/ManualFiles/manualE8110.pdf

Category: Peptide Protocols: Phage Display Cloning Systems and Protocols

Ph.D.â„¢-12 Phage Display Peptide Library Kit NEB. Applications for Drug Discovery using Phage Display Libraries of Peptides. - [Read Ph.D.â„¢-12 Phage Display Peptide Library Kit]

Protocol Guide for the N. crassa Yeast Artificial Chromosome Library - http://www.fgsc.net/fgn41/yac1.html

Category: Fungi Protocols: Neurospora Protocols

Protocol guide for the N. crassa yeast artificial chromosome library. Includes: Chromosome Walking; Hybridization screening of the YAC library; YAC restriction mapping and contig building; Preparation of chromosomal DNA plugs of YAC clones; Partial restriction enzyme digestion of YAC DNA plugs; Using CHEF gel analysis to resolve YAC clones; Southern Hybridization; Isolation of terminal restriction fragments from cloned DNA inserts in YAC clones; etc. - [Read Protocol Guide for the N. crassa Yeast Artificial Chromosome Library]

Protocols for Insect Cell Culture - http://www.hyclone.com/library/insectprotocols.htm

Category: Cell Culture Protocols: Insect Cell Culture Protocols

Includes protocols: Direct Adaptation to HyQ© SFX-Insect™ Medium; Sequential Adaptation to HyQ© SFX-Insect™ Medium; Adaptation of High Five™ Cells to HyQ© SFX-Insect™; Production of Recombinant Protein Using the Baculovirus Expression Vector System; Culturing Insect Cells and Production of Baculovirus Expressed Proteins in 2.8 L Fernbach Systems. Production of Autographa californica Multiple Capsid Nuclear Polyhedrosis Virus (rAcMNPV) Stock - [Read Protocols for Insect Cell Culture]

Rapid and efficient cDNA library screening by self-ligation of inverse PCR products (SLIP) - http://nar.oxfordjournals.org/cgi/reprint/33/21/e185?ijkey=XINgS7zgWnaIZMJ&keytype=ref

Category: RNA Protocols: cDNA Libraries Library

Hoskins et al. 2005 NAR. "... describe a PCR-based method for directed screening of plasmid cDNA libraries. " - [Read Rapid and efficient cDNA library screening by self-ligation of inverse PCR products (SLIP)]

RNA Isolation for cDNA Library Construction Protocol - http://www.oardc.ohio-state.edu/stockingerlab/Protocols/RNA%20Isolation.pdf

Category: DNA Protocols: cDNA Library Protocols

Protocol for RNA isolation for cDNA library construction. - [Read RNA Isolation for cDNA Library Construction Protocol]

Screening a cDNA Library for use with HybriZAP zebrafish cDNA libraries - http://www.cbs.umn.edu/~kclark/protocols/library.html

Category: RNA Protocols: cDNA Libraries Library

Procedures: Titer the library to determine the concentration of infectious phage, Plate and lift the phage particles, Hybridize the immobilized cDNA with labeled probe, Pick positive plaques. Karl Clark U. Minn. - [Read Screening a cDNA Library for use with HybriZAP zebrafish cDNA libraries]

Screening a Lambda Bacteriophage Genomic Library with a DNA Probe - http://omrf.ouhsc.edu/~frank/LIBSCRN.html

Category: RNA Protocols: cDNA Libraries Library

Frank Lab. Oklahoma Medical Research Foundation. How to screen a Lambda Bacteriophage Genomic Library with a DNA Probe. - [Read Screening a Lambda Bacteriophage Genomic Library with a DNA Probe]

Articles (1-3 of 3)

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.