Download e-book for iPad: Advanced Molecular Genetics by M. P. O’Connell, U. Priefer, N. Datta, M. E. Nugent (auth.),

By M. P. O’Connell, U. Priefer, N. Datta, M. E. Nugent (auth.), Alfred Pühler, Kenneth N. Timmis (eds.)

ISBN-10: 3642693059

ISBN-13: 9783642693052

ISBN-10: 3642693075

ISBN-13: 9783642693076

The improvement of robust new concepts and refmements of tech­ niques in molecular genetics lately, and the surge in curiosity in biotechnology in response to genetic equipment, have heralded a brand new golden age in molecular genetics, and motivated in varied disciplines a lot curiosity within the applied sciences themselves and their power makes use of in uncomplicated and utilized biomedical sciences. even supposing a few very good professional laboratory manuals (especially the chilly Spring Harbor Laboratory manuals by means of I. H. Miller; R. W. Davies et al. ; and T. Maniatis et al. ) on convinced chapters of molecular genetics exist, no normal textual content that covers a vast spectrum of the sub­ ject has so far been released. the aim of this handbook is to pre­ despatched such a lot, even though of necessity now not the entire vital tools of molecular genetics, in a chain of easy experiments, lots of which are effectively finished via the microbiologist, biochemist or biotechnologist that has had basically restricted publicity to genetics. the rest of the experiments require both higher familiarity with the topic, or assistance by means of anyone with such adventure. The booklet may still, consequently, not just let members to obtain new proce­ dures for ongoing tasks, but in addition function a foundation for the instructing of molecular genetic ideas in formal predoctoral and postdoctoral laboratory courses.

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Sample text

PRIEFER 1 Contents 1. General Introduction. . . . . . . . . . . . . . . . . . . 2. Experiment 1: Rapid Sizing of Intact Plasmid Molecules on Agarose Gels. . . . a) Introduction . . . . . . . . . . . . . . . . . . . . . b) Objectives . . . . . . . . . . . . . . . . . . . . . c) Procedure and Results . . . . . . . . . . . . . . . . . . 3. Experiment 2: Molecular Weight Determination of Restriction Fragments . . . .

3. Experiment 2: Conjugative Transfer of Plasmids to E. coli KI2. . . . . . a) Introduction. . . . . . . . . . . . . . . . . . . b) Objective. . . . . . . . . . . . . . . . . . . . c) Procedure . . . . . . . . . . . . . . . . . . . . . d) Materials. . . . . . . . . . . . . . . . . . . . . .. 4. Experiment 3: Transfer of Plasrnids to E. coli KI2 by Transformation . . .

In this section, only transformation N. E. Nugent 46 experiments involving restrictionless E. coli strains, such as E. coli C and E. f. are considered. Highest transformation frequencies are obtained using freshly isolated plasmid DNA that has been purified through caesium chloride-ethidium bromide (CsCI-EtBr) gradients and that contains a low proportion of nicked or broken molecules. However, DNA freshly isolated by other methods, such as that of Meyers et a1. (1976), may be used if very pure DNA is not available (for a review of different isolation methods see this volume).

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Advanced Molecular Genetics by M. P. O’Connell, U. Priefer, N. Datta, M. E. Nugent (auth.), Alfred Pühler, Kenneth N. Timmis (eds.)


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