Titlebook: Basic Techniques in Molecular Biology; Stefan Surzycki Book 2000 Springer-Verlag Berlin Heidelberg 2000 DNA.DNA Isolierung.DNA sequencing.

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Preparation of Genomic DNA from Plant Cells, from animal or bacterial cells. Plant cells have a very tough cell wall, the breakage of which requires the application of vigorous physical force that can shear large DNA molecules. Also, plant cells accumulate large amounts of sencondarymetabolities in their vacuoles that either co-purify with nu

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Isolation of Plasmid DNA, In most instances, plasmid DNA, isolated by these procedures, is of sufficient quality and quantity for cloning and PCR analysis. However, the plasmid DNA obtained by many of these methods is a poor template for double-stranded dideoxy DNA sequencing, largely due to inconsistency of template purity

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Isolation and Purification of RNA,, intact RNA is one of the central techniques in molecular biology and represents an important step in Northern analysis, nuclease protection assays, RNA mapping, RT-PCR, cDNA library construction and in vitro translation experiments.

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Isolation of PolyA+ RNA,emaining being ribosomal RNA and various species ofsmall stable RNAs. For some applications, such as construction of cDNA libraries or analysis of low-abundance mRNAs, isolation of mRNA instead oftotal cellular RNA is essential.

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Agarose Gel Electrophoresis of DNA,influencing the optimal separation of DNA bands in agarose gels. Protocols for running standard agarose gels and high resolution agarose gels are given, as well as a method for recovery of DNA fragments from the gel.

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Preparation of Probes for Hybridization,t one or both ends of nucleic acid molecules, giving specific, low density labeled probes. High density labeling is usually achieved by incorporating the reporters uniformly throughout entire length of nucleic acid molecules. For hybridization work internally labeled probes are preferred since they

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DNA Transfer and Hybridization, technique permits hybridization of various probes to immobilized target DNA under controlled conditions, coupled with fast detection of the hybrid. Because of the sensitivity, speed, and convenience of this “solid state” hybridization procedure, it has been widely applied in applied and basic resea