By Professor Dr. Volker Neuhoff (auth.)
This e-book is predicated on sensible event and is accordingly written as a prac tical guide. The fore-runners of the e-book have been the manuals of the 1st and moment EMBO-Courses on "Micromethods in Molecular Biology" which have been held in G6ttingen within the spring of 1970 and the fall of 1971. This ebook could function a guide not just for the members of the 3rd EMBO-Course to be held in G6ttingen in autumn 1973, but additionally for all experimenters who're drawn to utilizing micromethods. It has to be emphasised from the outset that this booklet is conceived as a "cook e-book" and never as a monograph which makes an attempt to revue the literature on micromethods seriously. The equipment defined the following intimately are played many times within the authors' laboratories and comprise the entire sensible info worthwhile for the winning appli cation of the micromethods. there are lots of different delicate and ideal micro equipment which aren't integrated during this ebook, as the authors consider that during a "cook ebook" in simple terms tools for which they've got own event and profi ciency may be defined. a few readers might think that the identify gives you greater than the current contents of this e-book; notwithstanding, if enough curiosity is proven during this quantity, it can be attainable to treatment such deficiencies in destiny editions.
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Additional info for Micromethods in Molecular Biology
After the glycerol has frozen completely, it is thawed out at room temperature, the gel slices are removed, transferred to the elution capillary and eluted as described. The glycerine adhering to the slices has no adverse effect on elution. Subsequent staining of the slices after elution demonstrates that it is not always possible to obtain complete elution, even in this way. Experiments with RNApolymerase from E. coli (see p. 28) show that the enzyme does not lose its enzymatic activity during elution by this method.
At 40 C in micro-dialysis chambers (see chapter 12) to remove the ammonium sulphate from the Glc-6-P DH suspension. If very dilute solutions are used, the stock solution need not be dialysed. 0 (1: lOw/v), and centrifuged at 00 C for 2 hrs. at 24000 g. The clear supernatant is divided into several portions and stored at - 200 C. 1 mg NADP/ml, is made just prior to electrophoresis. g. Glc-6-P DR, must be determined individually. 0). This amount is sufficient for the incubation of 20-40 microgels.
1g wet weights of rat kidney cortex. The dotted line represents the standard deviation of observations. 4 I-1g, respectively, were subjected to electrophoresis until the fluorescein marker had migrated 12 mm into the separation gel. The gels were then incubated for 15 min at 37° C. Fig. 35 demonstrates that the human and rat Glc-6-P DR pattern differs not only in the R f values, but also in the activity of the enzyme bands. On the right side of Fig. 35 the peak areas are plotted against I-1g wet weight of tissue.