control Protocols

Category: Cytogenetics Protocols: In Situ Hybridization Protocols

Protocol for the quality control of chromosome preparations. - [Read Quality Control of Chromosome Preparations Protocol]

Category: PCR Protocols: Reverse Transcriptase RT-PCR Protocols

In the first part of this protocol, the linear range of amplification is determined by carrying out 10 identical PCRs in the presence of [{alpha}-32P]dCTP and stopping one reaction after every two cycles.  Amplification products are quantified on a denaturing polyacrylamide gel and the results plotted on a graph (counts per minute vs. cycle number).  Total RNA is used as an internal control. - [Read Relative RT-PCR: Determining the Linear Range of Amplification and Optimizing the Primers:Competimer]

Category: PCR Protocols: Colony PCR

The following colony PCR protocol has been designed to be performed in individual reaction tubes. We usually test three colonies from each transformation along with a wild-type control. A series of five different PCR tests are performed on each colony to - [Read Single tube confirmation PCR protocol]

Category: Cell Biology and Cell Culture: In Vitro Reconstitution Protocols

Information for protocol using single-tube, coupled transcription/translation reactions for eukaryotic in vitro translation. Includes information on: Translation Procedure; Positive Control Translation Reactions Using Luciferase; Cotranslational Processing Using Canine Pancreatic Microsomal Membranes; Post-Translational Analysis; Positive Control Luciferase Assays; Composition of Buffers and Solutions; Luciferase SP6/T7 Control DNAs - [Read Single-tube Coupled Transcription/Translation Reactions for Eukaryotic In Vitro]

Category: RNAi Technology Protocols: RNAi Avian Embryos Protocols

Protocol describes a method to stain nerve fibers in tissue slices of avian embryos using an antibody against the 160-kD subunit of neurofilaments.  This allows the comparison of the branching pattern of motor and sensory neurons between control and experimental embryos.  The tissue is cut in slices using a vibratome or tissue slicer.  The protocol is suitable for older embryos after approximately stage 33 and regions that are not accessible by whole-mount analysis. - [Read Staining of Tissue Slices for Analysis of Axonal Pathfinding in dsRNA-Treated Avian Embryos]

Category: Avian Bird Protocols

Protocol describes a method to stain nerve fibers in tissue slices of avian embryos using an antibody against the 160-kD subunit of neurofilaments. This allows the comparison of the branching pattern of motor and sensory neurons between control and experimental embryos. The tissue is cut in slices using a vibratome or tissue slicer. The protocol is suitable for older embryos after approximately stage 33 and regions that are not accessible by whole-mount analysis. - [Read Staining of Tissue Slices for Analysis of Axonal Pathfinding in dsRNA-Treated Avian Embryos Protocol]

Category: Pharmacology and Toxicology Protocols: Cytotoxicity Protocols: Microorganism Cytotoxicity Assays Protocols

The basis of this test is that a cytotoxic chemical (regardless of site or mechanism of action) will interfere with the normal motility of the protozoan, Tetrahymena thermophila, in culture.  The degree of interference of motility as compared to control cultures, related to the concentration of the test compound, provides an indication of toxicity. - [Read Tetrahymena Thermophila Ocular Irritancy Test]

Category: Yeast Protocols: Yeast Two-Hybrid Systems Protocols

Protocol describes how to generate a plasmid construct (pBAIT) that expresses a target protein fused to the bacterial LexA protein. PBAIT is cotransformed into yeast with a lexAop-lacZ reporter plasmid carrying the bacterial lacZ gene under the control of the lexA operator. The recipient yeast strain contains a chromosomally integrated leu2 reporter gene, also under the control of the lexA operator. - [Read Two-hybrid Systems Stage 1: Characterization of a Bait-LexA Fusion Protein Protocol]

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]

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]

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]

Category: Neuroscience Protocols: Patch Clamping Protocols

The technique of patch clamp recording was invented by Bert Sakmann and Erwin Neher in 1981 for which they received the NOBEL prize. The technique is best suited for the study of the behaviour of single ion channels, or macroscopic currents in small cells or macro-patches. The whole cell technique allows one to control the composition of solutes on both sides of the membrane. - [Read Whole Cell Patch Clamp Technique]

Category: RNAi Technology Protocols: RNAi Avian Embryos Protocols

Protocol describes a method for staining nerve fibers in whole-mount preparations of avian embryos using an antibody against the 160-kD subunit of neurofilaments.  This allows the comparison of the branching pattern of motor and sensory neurons between control and experimental embryos.  This protocol has been successfully applied for embryos at different stages up to about stage 33 (7 days of incubation). - [Read Whole-Mount Preparations for Analysis of Axonal Pathfinding in dsRNA-Treated Avian Embryos]

Category: Avian Bird Protocols

Protocol describes a method for staining nerve fibers in whole-mount preparations of avian embryos using an antibody against the 160-kD subunit of neurofilaments. This allows the comparison of the branching pattern of motor and sensory neurons between control and experimental embryos. This protocol has been successfully applied for embryos at different stages up to about stage 33 (7 days of incubation). - [Read Whole-Mount Preparations for Analysis of Axonal Pathfinding in dsRNA-Treated Avian Embryos Protocol]