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Screening an Unamplified Cosmid Library by Hybridization: Plating the Library onto Filters Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3289

Category: DNA Protocols: Cosmid Library Protocols

Unamplified cosmid libraries are plated at high density onto 150-mm nitrocellulose or nylon filters and screened by hybridization - [Read Screening an Unamplified Cosmid Library by Hybridization: Plating the Library onto Filters Protocol]

Screening Expression Libraries Constructed in Bacteriophage Lambda Vectors Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3765

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

An expression library constructed in a bacteriophage {lambda} vector is plated on an appropriate E. coli strain in the absence of isopropylthio-ß-D-galactoside (IPTG). After 2-4 hours, the plates are moved to 37°C (to stabilize any fusion proteins that are temperature sensitive), and filters impregnated with IPTG are laid on top of the developing plaques. - [Read Screening Expression Libraries Constructed in Bacteriophage Lambda Vectors Protocol]

Screening Expression Libraries Constructed in Plasmid Vectors Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot4079

Category: DNA Protocols: cDNA Library Protocols

A cDNA library constructed in a plasmid expression vector of the pUC, pUR, or pEX series is plated on agar medium and then replicated onto filters, which are transferred to plates containing IPTG. After 2-4 hours of induction, the colonies are lysed with chloroform and then screened with appropriate antibodies. - [Read Screening Expression Libraries Constructed in Plasmid Vectors Protocol]

Shotgun Library Construction Protocol - http://www.oardc.ohio-state.edu/stockingerlab/Protocols/ShotgunLibraryConstruction%28EJSVersion-050511%29.pdf

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

Protocol for shotgun library construction. - [Read Shotgun Library Construction Protocol]

Staining for Myelin Sheath Protocol - http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/LFB.PDF

Category: Neuroscience Protocols: Neuronal Stains Protocols

Protocol for myelin sheath. Luxol Fast Blue is the alcohol soluble counterpart of the water soluble Alcian Blue. Staining is due to lipoproteins, and the mechanism is one of an acid-base reaction with salt formation; the base of the lipoprotein replaces the base of the dye. - [Read Staining for Myelin Sheath Protocol]

Staining of Nerve Fibers Protocol - http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/BODIAN.PDF

Category: Neuroscience Protocols: Neuronal Stains Protocols

Protargol-S (silver proteinate) is used with the addition of copper metal. The copper replaces the silver in the connective tissue, allowing a greater differentiation between the nerve fibers and the connective tissue. The silver is reduced with hydroquinone to the visible metallic form. The sections are toned with gold chloride, the gold chloride is reduced with oxalic acid, increasing the deposit of metallic gold on the sections. - [Read Staining of Nerve Fibers Protocol]

Synthesis of DNA Libraries: Use of PCR to Analyze Ligation Substrates and Products - http://www.cshprotocols.org/cgi/content/extract/2006/14/pdb.prot4136

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

A vector that lacks inserts can significantly contaminate a DNA library. This contamination can occur either from self-ligation of single-cut vector or from uncut circular starting plasmid. - [Read Synthesis of DNA Libraries: Use of PCR to Analyze Ligation Substrates and Products]

Synthesis of Radiolabeled, Subtracted cDNA Probes Using Oligo(dT) as a Primer Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3869

Category: PCR Protocols: cDNA Synthesis Protocols

Protocol describes the preparation of subtracted cDNA probes by hybridization to an mRNA driver, followed by purification of the single-stranded radiolabeled cDNA by hydroxyapatite chromatography. Before preparing the probe, it is a good idea to have filters (which contain the cDNA library to be screened) ready to hybridize. - [Read Synthesis of Radiolabeled, Subtracted cDNA Probes Using Oligo(dT) as a Primer Protocol]

Systematic screening of an arrayed cDNA library by PCR PDF - http://www.pnas.org/cgi/reprint/92/6/2209

Category: RNA Protocols: cDNA Libraries Library

Munroe et al PNAS 1995 "..developed a PCR-based method for rapid and effective screening of arrayed cDNA libraries. - [Read Systematic screening of an arrayed cDNA library by PCR PDF]

Thioflavine S Amyloid Protocol - http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/THIOFLAV.PDF

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

Protocol for thioflavine amyloid. - [Read Thioflavine S Amyloid 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]

Two-hybrid Systems Stage 2: Selecting an Interactor Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3888

Category: Yeast Protocols: Yeast Two-Hybrid Systems Protocols

In this stage of the protocol, a mammalian cDNA library constructed in a plasmid such as pJG4-5 is transformed into yeast strains containing pBAIT and the lexAop-lacZ reporter plasmid. PJG4-5 expresses the cloned cDNAs from a cassette containing a transcriptional activation domain and other moieties under the control of the yeast GAL1 promoter. - [Read Two-hybrid Systems Stage 2: Selecting an Interactor Protocol]

Use of PCR for Quality Control of a Genomic DNA Library Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4138

Category: DNA Protocols: Genomic DNA Library Protocols

Protocol describes the use of PCR to analyze the quality of a human genomic DNA library. It has been optimized for a human genomic DNA library containing fragments between approximately 50 and 500 bp. - [Read Use of PCR for Quality Control of a Genomic DNA Library Protocol]

Use of PCR for Quality Control of a Peptide DNA Library Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4137

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

Protocol describes the use of PCR to characterize a peptide library encoded in a plasmid vector. In this example, the library was obtained by transforming bacteria with the ligation reaction at the end of Use of PCR to Prepare a Double-Stranded DNA Library Encoding Random Peptides. - [Read Use of PCR for Quality Control of a Peptide DNA Library Protocol]

Use of PCR to Prepare a Double-Stranded DNA Library Encoding Random Peptides Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/14/pdb.prot4135

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

PCR is used as a preparative tool for the synthesis of a high-complexity double-stranded DNA library. In the example presented here, a mixture of synthetic oligonucleotides is used to synthesize a random peptide NNK library, where K is either T or G. The exclusion of A and C nucleotides at the third position decreases the occurrence of stop codons but still allows codons for all 20 amino acids. - [Read Use of PCR to Prepare a Double-Stranded DNA Library Encoding Random Peptides Protocol]

Yeast Transformation (for LexA Library Screen) Protocol - http://www.natureprotocols.com/2006/12/15/yeast_transformation_for_lexa.php

Category: Yeast Protocols: Yeast Transformation Protocols

Protocol for Yeast transformation (for LexA Library Screen). - [Read Yeast Transformation (for LexA Library Screen) Protocol]

Yeast Two-Hybrid Screen with Library and Bait - http://faculty.virginia.edu/mirmira/resources_files/Protocols/YEAST%20TWO.htm

Category: Molecular Model Organisms: Yeast Two-Hybrid Screen Protocol

yeast transformed with bait vector. yeast culture. selection plates. Mirmira, Univ. Virginia. - [Read Yeast Two-Hybrid Screen with Library and Bait]

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.