Nucleic acid isolation

An alternative to nucleic acid isolation by electrophoresis followed by electrophoretic elution is HPLC. There are several advantages in the use of HPLC. It is simple and easy to use and has a high degree of resolution, it is faster than electrophoresis and contamination of DNA fragments by impurities from the agarose gel is avoided. 

The discovery of deoxyribonucleic acid dates to 1869, when Miescher isolated a new chemical substance from white blood cells that he obtained from pus and later from sperm cells.3 The material, which became known as nucleic acid, occurred in both plants and animals, thymus glands and yeast cells being among the best sources. Chemical studies indicated that the nucleic acids isolated from thymus glands and from yeast cells were different. As we now know, thymus nucleic acid was primarily DNA and yeast nucleic acid primarily RNA. For a while it was suspected that animals contained only DNA and plants only RNA, and it was not until the early 1940s that it was established that both substances were present in all organisms.3 ... 

As with the first two editions, the book is organized into two parts I. Theory and Experimental Techniques and II. Experiments. Part I introduces students to theoretical and background material for the experiments. This part may also serve as a supplement for instructors who use their own experiments. In Part II there are 15 experiments that represent all areas of biochemistry, including working with proteins and nucleic acid isolation and characterization. The number of experiments has been reduced from earlier editions at the request of instructors and students who believed the book had more experiments than needed for a typical one-semester course. There are, however, still sufficient experiments for a two-semester course sequence. The reduction in the number of experiments has also been achieved by combining some experiments. 

Desoxy-D-ribose occurs naturally as the carbohydrate component of desoxyribonucleic acids. From an examination by the ultraviolet chromatographic technique, of a range of nucleic acids isolated from animal, plant and bacterial sources, Chargaff and his coworkers10 have concluded that this sugar is the only 2-desoxypentose found so far to occur naturally in the nucleic acids. 

Nowadays the genomics-based techniques mainly employed include microarrays [31], serial analysis of gene expression (SAGE) [32] and circulating nucleic acids isolated from plasma or serum (CNAPS) [33]. 

C. Circulating Nucleic Acids Isolated from Plasma or Serum (CNAPS)... 

The complexity in morphological form and internal cellular organization among algal representatives (Table I) requires that separate protocols, unique from those methodologies devised for terrestrial plant nucleic acid isolation, be developed. This chapter presents several protocols for the extraction and isolation of algal nucleic acids. 

General aspects of protein and nucleic acid isolation... 

A nucleic acid isolation area, where new samples are stored and submitted to the extraction procedure. 

The MagNA Pure LC Instrument allows automated isolation and preparation of nucleic acids. The instrument performs up to 32 nucleic acid isolations in 1 to 3 hours from various specimens, which can include blood, blood cells, culture cells, plasma, serum, sputum, stool, broncho-alveolar lavage, plant tissues, or food products. 

Principles of Molecular Biology and Approaches to Nucleic Acid Isolation... 

Direct PCR product analysis by electrophoresis is also frequently used in the clinical laboratory to query the quality of intermediary steps prior to the assay. (For example, was the nucleic acid isolated successfully How well was it purified Did the amplification work How specific was the PCR And so on). 

Information with regal d to specimen types, specimen handling and transport procedures, nucleic acid isolation and storage, description of the test procedure, data reports, expected results, technical interpretation of results. 

Nivens D, Applegate B, Method for nucleic acid isolation using supercritical fluids. US patent 5922536, 1999. 

An integrated pTAS system for the detection of bacteria including lysis, DNA purification, PCR and fluorescence readout has also been published recently [113]. A microfluidic plastic chip with integrated porous pol5mier monoliths and silica particles for lysis and nucleic acid isolation was used for detection (Fig. 8). A custom-made base device provided liquid actuation and off-chip valving by stopping liquid flow from the exits of the chip, utilizing the incompressibility of liquids. Detection of 1.25 x 10 cells of B. subtilis was demonstrated with all assay steps performed on-chip. 

Uridine 5 -Diphosphate. Uridine 5. (trihydrogen diphosphate) UDP uridine 5 -pyrophosphate uridine-5-pyrophosphoric acid. C9HI4N20,jPj mol wt 404.18. C 26.75%, H 3.49%, N 6.93%, O 47.50%, P 15.33%. Can be isolated from calf s fiver, thymus, and yeast. The commer -cial product is derived from yeast. Pentose nucleic acids (isolated from yeast) are digested with rattlesnake venom (freed of 5 -monoesterase) and the nucleotides are separated by chromatography Cohn, Volkin, Arch, Biochem. Biophys. 35, 465 (1952) J. Biol Chem. 203, 319 (19S3). For alternate procedures see the refs under Uridine Diphosphate Glucose. Syntheses Chambers, J. Am. Chem. Soc. 81, 3032 (1959) Moffatt, Khorana, ibid. 83, 649 (1961). 

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