alkaline Protocols

Category: Nucleic Acid Electrophoresis Protocols: DNA Electrophoresis Protocols: DNA Electrophoresis in Agarose Gels Protocols

Alkaline agarose gels are used chiefly to measure the size of first and second strands of cDNA (Construction of cDNA Libraries Stage 1: Synthesis of First-strand cDNA Catalyzed by Reverse Transcriptase) and to analyze the size of the DNA strand after digestion of DNA-RNA hybrids with nucleases such as S1. - [Read Alkaline Agarose Gel Electrophoresis Protocol]

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

Protocol for alkaline congo red technique. - [Read Alkaline Congo Red Technique Protocol]

Category: Microbiology: Mini-Prep Plasmid Purification

Harvesting and Lysis of Bacteria. Mini-prep plasmid purification using Solution I, Solution II and Solution III. The Preuss Lab. Univ. Chicago. - [Read Alkaline Lysis Mini-Prep Protocol]

Category: Signalling Signal Transduction Protocols

Technique for Alkaline Phosphatase (APAAP). Techniques includes information on: Cytological Preparations, Fixation, Technique, Results, Substrate Solution and 0.005M Tris/HC1 Saline Buffer pH 7.6. - [Read Alkaline Phosphatase (APAAP) Technique]

Category: Immunohistochemistry Protocols: Antibody Staining Protocols

Protocol for alkaline phosphatase (APAAP) technique. Includes: Fixation; Technique; Substrate Solution. - [Read Alkaline Phosphatase (APAAP) Technique Protocol]

Category: Neuroscience Protocols: Histology Stains Protocols: Enzyme Histochemical Stains Protocols

Protocol for alkaline phosphatase histochemical stain. - [Read Alkaline Phosphatase Histochemical Stain Protocol]

Category: Viral Manipulation Protocols: Phage Lamda Manipulation Protocols

Removal of the 5'-phosphate groups from the internal termini of bacteriophage {lambda} arms is used to prevent self-ligation and suppress the background of nonrecombinant bacteriophages during cloning. - [Read Alkaline Phosphatase Treatment of Bacteriophage lambda Vector DNA Protocol]

Category: Microbiology: Bacterial E.Coli Culture Media & Plates

agar, Ampicillin Stock, Alkaline Lysis Solution, SOB, SOC media, and X-gal stock solutions. Donis Keller. - [Read Bacterial Media Solutions and Stocks]

Category: Protein Protocols: Protein Quantitation Concentration Protocols: Bradford Protein Assay Protocols

The bradford dye-binding assay is a colorimetric assay for measuring total protein concentration.  It involves the binding of Coomassie Brilliant blue to protein. There is no interference from cations nor from carbohydrates such as sucrose. 
Detergents such as sodium dodecyl sulfate and triton x-100 can interfere with the assay, as well as strongly alkaline solutions.

Includes a general overview of the procedure and preparation of the standards in the protocol. - [Read Bradford Assay Method]

Category: Buffers Media, and Solutions: Buffer Theory

This page describes simple acidic and alkaline buffer solutions and explains how they work. - [Read Buffer Solutions and How they work...]

Category: Viral Manipulation Protocols: M13 Phage Manipulation Protocols

Protocol describes three standard methods to construct bacteriophage M13 recombinants: (1) ligating insert DNA to a linearized vector, prepared by cleavage of M13 RF with a single restriction enzyme; (2) using alkaline phosphatase to suppress self-ligation of the linearized vector, and (3) using M13 RF cleaved with two restriction enzymes for directional cloning. - [Read Cloning into Bacteriophage M13 Vectors Protocol]

Category: Neuroscience Protocols: Histology Stains Protocols: Enzyme Histochemical Stains Protocols

Protocol for detection of ß-Galactosidase and alkaline phosphastase activities in tissue. - [Read Detection of ß-Galactosidase and Alkaline Phosphastase Activities in Tissue Protocol]

Category: Cytogenetics Protocols: DIG Biotin Labeled Probe Protocols

Protocol describes here a high sensitivity indirect detection procedure for DIG-labeled hybridization probes. The procedure uses the components of the HNPP Fluorescent Detection Set to form a fluorescent precipitate of HNPP (2-hydroxy-3-naphthoic acid-2’-phenylanilide phosphate) and Fast Red TR at the site of hybridization. Includes: In situ hybridization with DIG-labeled probes; Detection of DIG-labeled probes; Fluorescence microscopy. - [Read DNA In Situ Hybridization with an Alkaline Phosphatase-Based Fluorescent Detection System]

Category: Nucleic Acid Modification Protocols: DIG Biotin Labeled Probes Protocols

Protocol describes a high sensitivity indirect detection procedure for DIG-labeled hybridization probes.  The procedure uses the components of the HNPP Fluorescent Detection Set to form a fluorescent precipitate of HNPP (2-hydroxy-3-naphthoic acid-2’-phenylanilide phosphate) and Fast Red TR at the site of hybridization.  This procedure can be used to detect single copy sequences as small as 1 kb on human metaphase chromosomes. 
    - [Read DNA In Situ Hybridization with an Alkaline Phosphatase-Based Fluorescent Detection System Protocol]

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]

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]

Category: Nucleic Acid Purification Protocols: DNA Isolation Protocols: DNA Plasmid Preps Protocols

Protocol for DNA preparation, plasmid DNA using alkaline method. - [Read DNA Preparation, Plasmid DNA Alkaline Method Protocol]

Category: Immunohistochemistry Protocols: Antibody Staining Protocols

Protocol for histochemical staining. This protocol has been optimized for b-galactosidase and human placental alkaline phosphatase staining in retinal tissue and cultured cells. - [Read Histochemical Staining Protocol]

Category: Immunohistochemistry Protocols

Protocol describes the application of peroxidase or alkaline phosphatase conjugates in the immunohistochemical labeling of formalin-fixed, paraffin-embedded tissue sections. Includes: Removal of Paraffin and Rehydration; Antigen Retrieval - Unmasking of Antigen; Enzyme retrieval; Microwave retrieval; Inactivation of Endogenous Peroxidase; etc.. - [Read Immunohistochemistry Protocol]

Category: BACs Bacterial Artificial Chromosome Protocols

BAC DNAs are prepared from 5-ml cultures of BAC-transformed cells by a modification of the standard alkaline lysis method (Preparation of Plasmid DNA by Alkaline Lysis with SDS: Minipreparation). The yield typically varies between 0.1 and 0.4 µg of BAC DNA. - [Read Isolation of BAC DNA from Small-scale Cultures Protocol]

Category: Protein Protocols: Lowry Protein Assay

Lowry Protein Assay. The Lowry procedure is one of the most venerable and widely-used protein assays, being first described in 1951 [Lowry et al., J. Biol. Chem. 193: 265-275 (1951)]. Under alkaline conditions, copper complexes with protein. When folin phenol reagent (phospho-molybdic-phosphotungstic reagent) is added, the Folin-phenol reagent binds to the protein. Bound reagent is slowly reduced and changes color from yellow to blue. P.J. Hansen, Dept. of Animal Sciences, University of Florida. - [Read Lowry Protein Assay]

Category: Mouse Protocols: Mouse Reproductive System Surgery Protocols

Protocol describes how to remove the gonads, determine the sex ofmouse embryos 13.5 days post coitum and visualize the germ cells, through use of an alkaline phosphatase staining. - [Read Mouse Gonad Removal and Germ Cell Staining Protocol]

Category: DNA Protocols: SAP Protocols

PCR cleanup using shrimp alkaline phosphotase (SAP). Orti Lab. - [Read PCR cleanup using shrimp alkaline phosphotase (SAP)]

Category: DNA Protocols: SAP Protocols

Phosphatasing with Shrimp Alkaline Phosphatase (S.A.P.). Shirley Reynolds Hendrick Lab. - [Read Phosphatasing with Shrimp Alkaline Phosphatase (S.A.P.)]

Category: Chromatography Protocols: Dye Ligand Affinity Chromatography Protocols

Protocol describes a general approach for the direct attachment of a triazine dye onto a carbohydrate support.  The direct coupling method is achieved under alkaline conditions (pH 10-11) with nucleophilic displacement of the dye’s chlorine atom by the polymer hydroxyls. - [Read Preparation of Affinity-Ligand Resins by Immobilization of Dyes on Polyhydroxyl Matrices Protocol]