A simplified method for determining the P50 of blood.

A method to determine the P50 of whole blood is described using a modified American Optical reflectance oximeter, pump, and membrane tonometer, together with PO2, PCO2, and pH measurements in a standard blood gas machine. Determinations of P50 were made in 66 patients and normal subjects and in two situations where P50 was very low and very high. The results were compared to oxygen saturations calculated from measured oxygen content. The directly determined oxygen saturation agreed with the assumed saturation of 50 per cent in the oximeter within 0.5 per cent. The apparatus appears to be a simple and relatively inexpensive method to obtain P50 as long as blood carboxyhemoglobin or methemoglobin contents are not elevated.     

A computer program for determining the size of DNA restriction fragments

A computer program has been developed for determining the sizes of DNA restriction fragments from their electrophoretic mobilities. A parabola is fitted to the mobilities of a set of standard fragments of known sizes and the sizes of the unknown fragments are then calculated from the fitted curve. This procedure is shown to yield estimated sizes which are accurate to within a few percentage, as judged by experiments with fragments obtained by digestion of pBR322 with the restriction endonuclease HaeIII. The program, which is written in BASIC, is simple to use and is very much faster than the graphical method that it replaces.     

Direct DNA sequencing of PCR amplified genomic DNA by the Maxam-Gilbert method

We present a method to directly sequence PCR amplification products utilizing the chemical method of Maxam and Gilbert. This procedure yields clearly readable DNA sequences of 100-400 base pairs in length derived from human genomic DNA in four days' time.     

A simplified routine method for determining carbohydrate fermentation by yeasts

The use of commercially available materials in a minimal medium contributes to the simplicity, reliability, and reproducibility of a method described to demonstrate carbohydrate fermentation reactions by yeasts. The medium consists of 1% yeast extract and 2% test carbohydrate in distilled water, dispensed in a modified Durham fermentation tube. Carbohydrate fermentation patterns can usually be obtained within a period of 7 days. The ability of yeasts to ferment carbohydrates is determined strictly on the basis of gas production from the substrate. The method proved reliable in reproducing established fermentation patterns for 112 different yeast strains representing 13 separate genera.

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Zusammenfassung Der Gebrauch von handelsüblichen Grundbestandteilen in einem geringen Medium trägt zu der Einfachheit, Zuverlässigkeit und Reproduzierbarkeit der beschriebenen Methode, um die Gärungsreaktionen der Karbohydrate durch Hefen zu veranschaulichen, bei. Das Medium besteht aus einem 1 prozent. Hefenauszug und aus einem 2 prozentigen Testkarbohydrat in destilliertem Wasser, die auf modifiziertes Durham Gärungsreagensglas verteilt sind. Das Karbohydrat-Gärungsmodell kann gewöhnlich innerhalb einer Periode von sieben Tagen erhalten werden. Die Fähigkeit von Hefen, Karbohydrate zu vergären, ist ausschließlich auf Grund der Gasentwicklung aus den Produkten bestimmt. Die Methode erwies sich zuverlässig in der Erzeugung festgestellter Gärungsmodelle aus 112 Hefen, die 13 verschiedene Gattungen darstellten.

     

A hybrid symbolic-numerical method for determining model structure

In this article, we present a method for determining whether a model is at least locally identifiable and in the case of non-identifiable models whether any of the parameters are individually at least locally identifiable. This method combines symbolic and numeric methods to create an algorithm that is extremely accurate compared to other numeric methods and computationally inexpensive. A series of generic computational steps are developed to create a method that is ideal for practitioners to use. The algorithm is compared to symbolic methods for two capture-recapture models and a compartment model.     

A simplified method of rapid hemagglutination test for determining the levels of tetanus antibodies]

A new, simplified methodology of preparation of lyophilized chrome-coated turkey blood cells (formalin-treated ) for rapid identification of tetanus antibodies in indirect hemagglutination test is described. Blood cells diagnostic preparations obtained in this way were easier and three times faster to prepare than tanned cell preparations . They maintained unchanged capability for specific agglutination during at least one year of storage when kept at 4 degrees C. proposed methodology enable to start a production of laboratory kits necessary for controlled prophylaxis of tetanus.     

Cloning and determining the primary structure of fragments of the RNA-polymerase beta-subunit genes from representatives of Baikal picoplankton

Fragments of the RNA polymerase beta-subunit genes from representatives of picoplankton of Lake Baikal were cloned using polymerase chain reaction (PCR), and the resulting recombinant clones were sequenced. An analysis of 93 clones revealed 40 different sequences. A comparison of these nucleotide sequences and the corresponding amino acid sequences deduced with their homologues from databanks confirmed the high conservation of the RNA polymerase beta-subunit region involved in the formation of the enzyme active site.     

A method of determining fragments of peptide bioregulators responsible for interaction with receptors

A method for investigating the relationship between chemical structure of peptide molecules and their biological activity is suggested. It is based on a few statistical algorithms which are described. The results of the method testing on the thyrotropin-releasing hormone analogs are presented.     

A method for determining DNA and chondrocyte content of articular cartilage

A novel and precise method was devised to study the DNA and chondrocyte content of articular cartilage. It involved the sequential digestion of cartilage matrix with hyaluronidase, trypsin, and collagenase to release the chondrocytes. A direct cell count and DNA assays were then performed on the cells. The concentration of cells was the quotient of the total number of cells and the weight of cartilage used. The DNA content of cartilage is identical to the amount of DNA in the chondrocytes. Our data also confirmed the earlier findings that cell density and DNA content of articular cartilage decreased gradually to a relatively constant level as animals matured to adulthood.     

A method for recovering strand-specific probes from nick-translated DNA fragments

A method of preparing strand-specific probes for DNA X DNA or DNA X RNA hybridizations is described. Double-stranded DNA fragments are first isolated from any recombinant DNA clone containing the desired sequence, and then labeled in vitro by nick-translation (T. Maniatis, A. Jeffrey, and D. G. Kleid (1975) Proc. Natl. Acad. Sci. USA 72, 1184-1188; P. W. J. Rigby, M. Dieckmann, C. Rhodes, and P. Berg (1977) J. Mol. Biol. 113, 237-251). Sequences homologous to the desired strand are captured by annealing the denatured nick-translate to viral strands of an appropriate M13 clone, and recovered by elution of the resulting hybrids from a column of agarose A50M (Bio-Rad). By this method, separate probes with specificity to either strand, as well as the double-stranded probe, may conveniently be prepared from a single nick-translation reaction. Probes may be obtained which are homologous either to the full length of the cloned region or to selected portions thereof by selecting appropriate M13 clones for annealing. The probe is recovered as a population of fragments several hundred bases or less in length, which have been found ideal for saturating liquid hybridizations, and should be similarly well suited for in situ hybridizations to cytological preparations.     

A general method for cloning DNA fragments in multiple copies

A general method for the cloning of DNA fragments in tandem arrays is presented. Synthetic directional adapters are attached to the fragment ends to establish complete control over fragment orientation during ligation. Use of these directional adapters thereby ensures the production of direct repeats, an arrangement essential for clone stability. The technique involves a protocol that is both less complex and less time-consuming than previous methods. Using this technique, a 10-min ligation has yielded a plasmid containing 20 copies of a 151-bp fragment.     

On the algorithms for determining the primary structure of biopolymers

The algorithm for determining the primary structure of biopolymers from complete and partial digests are analyzed. The problem of determining the primary structure is formulated in the form of the problem of word reconstruction in the limits of which the corresponding algorithms are analyzed. Difficulties arising in constructing the algorithms for determining the primary structure of nucleic acids from a partial digest are discussed. They seem to be due to the extensive testing of variants. When there is a certain scheme of the initial data from a partial digest we propose an economical testing (searching) algorithm. The scheme of an effective algorithm for reconstruction of the primary structure fromN complete digests is given.