accurate Protocols

Category: Chromatography Protocols: Liquid Chromatography Protocols

Protocol for configuration, column construction, and column packing for a capillary liquid chromatography system. Protocol describes a procedure for adapting conventional HPLC systems to provide accurate low-flow rates (0.4-4 µl/min) and gradients required to operate slurry-packed capillary columns.  A key component of this system is a commercial axial-beam longitudinal flow cell that can be fitted to several commercial UV detectors. - [Read Configuration Column Construction Column Packing for Capillary Liquid Chromatography]

Category: Cytogenetics Protocols: In Situ Hybridization Protocols

In situ methods to visualize transgenes (including single copy genes) and their transcripts during interphase from different tissues and plant species. These techniques reduce the time necessary for characterization of transgene integration by eliminating the need for time-consuming segregation analysis, and extend characterization to the interphase nucleus, thus increasing the likelihood of accurate prediction of transgene activity. - [Read In Situ Methods to Localize Transgenes and Transcripts in Interphase Nuclei]

Category: PCR Protocols

Protocol illustrates the rules of successful long PCR: No more than 1 ng of template DNA is used per microliter of PCR in a 100-µl reaction; approximately 0.1 µl of KlentaqLA (not plain Taq) is used per kilobase of target (for targets >10 kb, 1-1.3 µl of enzyme should be used); the Mg++ concentration is considered as the excess over the level of dNTPs. - [Read Long and Accurate PCR Protocol]

Category: Chromatography Protocols: Liquid Chromatography Protocols

Protocol describes a procedure for adapting conventional HPLC systems to provide accurate low-flow rates (0.4-4 µl/min) and gradients required to operate slurry-packed capillary columns.  A key component of this system is a commercial axial-beam longitudinal flow cell that can be fitted to a number of commercial UV detectors. - [Read Packing Capillary Columns for RP-HPLC Protocol]

Category: Bacteria Genetics Protocols

Protocol describes a method to determine the presence of plasmid DNA in an Agrobacterium culture. Compared to selection of transformed Agrobacterium, which can be ambiguous and normally takes several days for resistant colonies to appear, the approach described here is both rapid and accurate. - [Read PCR Analysis of Agrobacterium Protocol]

Category: Drosophila Protocols: Drosophila Genetics Protocols

Protocol describes how to produce a soluble nuclear extract rich in basal pol II transcription factors from Drosophila embryos. This is a cell-free extract that contains all the necessary transcription factors and is capable of accurate initiation of transcription by RNA polymerase II but is deficient in core histones and histone H1. - [Read Preparation of a Highly Efficient Transcription Extract from Drosophila Embryos Protocol]

Category: Signalling Signal Transduction Protocols

Detection of phosphorylated tyrosine residues can be performed using anti-P-TYR Ab and Western Analysis.Includes 2nd method,which uses phosphotyrosine in conjunction with anti-P-TYR Ab to "unlabel" potential proteins.By comparing Westerns developed with the 1st method(reveals phosphorylated protein) and the 2nd method(reveals non-specific labeling), a more accurate picture of those proteins phosphorylated on tyrosine can be seen. Includes: Protein Preparation, Electrophoresis and Transfer. - [Read Protocol for Antiphosphotyrosine Western Blot Analysis]

Category: Protein Protocols: Protein Precipitation Procols

A protein precipitation procedure used to precipitate protein from cell lysates. Allows for optimal protein recovery and accurate assays. The Signaling Gateway - [Read TCA/Acetone Precipitation (Large Scale) PDF]

Category: Western Blot Protocols

Procedure describes how it denatures most of the modification and degradation proteins immediately giving the most accurate read out of the true levels of protein at the time of harvest.  However, in cases where detection is a problem, a limited purification (e.g. isolation of nuclear extract for the detection of transcription factors) might be required to allow analysis. - [Read Western Blot Analysis of Endogenous Gene Expression Protocol]