Extraction of nucleic acid

Recently, the use of AR has extended into several other areas, yielding interesting information for cytology, fresh cell/tissue sections, and fluorescence IHC (fluorescence in situ hybridization [FISH]), in addition to adaptations of the method for extraction of nucleic acids and proteins from FFPE tissues for use with modern methods of molecular analysis. In this chapter, the emphasis is on expanded applications in diagnostic cytology, fresh frozen cell/... 

The first step in the extraction of nucleic acids may require the lysis of cells and the inactivation of cellular nucleases. These two steps may be combined into one. For instance, a single solution may contain detergents to solubilize the cell membrane and strong chaotropic salts to inactivate the intracellular... 

The earlier work on the isolation of PolyPs from the cells of living organisms usually employed the same methods as those used for the extraction of nucleic acids. It was not until 1936 that MacFarlane (MacFarlane, 1936) proposed a specific method for the extraction and fractionation of condensed phosphates present in cells. It was found that these phosphates could be divided into two main fractions, i.e. one soluble in 5 % trichloroacetic acid (TCA) and the other insoluble, and ever since then cellular condensed polyphosphates have been divided into acid-soluble and acid-insoluble fractions. 

This chapter illustrates the characterizations of technical procedures necessary in the course of these molecular evaluations, describing our laboratory experience and suggesting different diagnostic approaches microdissection techniques, extraction of nucleic acids from paraffin-embedded tissues, polymerase chain reaction (PCR), direct sequencing, and allelic separation by cloning [4-7],... 

One application example on the microfluidic LSI platform is the extraction of nucleic acids (NA) from a small amount of cells [126, 127] for cell-based assays. For the extraction of NA from a cell suspension, the cell membrane has to be destroyed first (chemical lysis of the cell). Afterwards, the NA are specifically separated from the residual cell components using a solid phase extraction method based on an NA affinity column (paramagnetic beads). This extraction protocol is completely implemented on the microfiuidic... 

Due to methodological problems it is presently not possible to quantifying those taxa which are determined to be present in clone libraries these problems, that are associated with the extraction of nucleic acids, PCR-primer sensitivity and selectivity, cloning steps, and the dependence of PCR amplificate amount on undeterminable genomic properties, have been summarized [21]. Thus, it is also not possible to decide whether the identified clones belong to a majority or a minority population of the naturally occurring prokaryotes. 

Reedy CR, Hagan KA, Strachan BC, Higginson JJ, Bienvenue JM, Greenspoon S A, Ferrance JP, Landers JP (2010) Dual-domain microchip-based process for volume reduction solid phase extraction of nucleic acids from dilute, large volume biological samples. Anal Chem 82 5669-5678... 

Bhattacharyya A, Klapperich CM (2005) Polymeric microfluidic device fOT on-chip cell lysis and extraction of nucleic acids frran biological samples. In Proceedings of the 9th intemational confertaice miniaturized systems chemistry life sciences, Boston, pp 1167-1169... 

Phenol-chloroform mixtures from extraction of nucleic acids from radiolabeled cell components. 

Extraction of nucleic acids from environmental samples accounts for a large fraction of the microorganisms that is not culturable but may be responsible for major biodegradation activities. 

The construction of a clone library involves the extraction of nucleic acids from environmental samples followed by PCR-amplification of the target gene... 

Fig. 7 Principle of the T-RFLP technique. Following extraction of nucleic acids, PGR amplification is performed using one fluorescently labeled primer. The amplified products are then subjected to restriction digestion, which leads to the generation of fluorescently labeled fragments of different lengths. The fragments are separated on a sequencing gel...

A main issue in culture-independent methods relates to the extraction of nucleic acids, because technical issues may arise DNA may not be recovered from all genotypes, which could remain undetected due to low species abundance, insufficient homogenization of the matrix, or inadequate cell lysis that prevents the release of nucleic acids. Moreover, even if DNA yield is high, macromolecule inhibitors that have not been eliminated may lower PCR sensitivity, and PCR amplification may be inaccurate or inhibited (Jany Barbier, 2008). For these reasons, protocols have to be accurately adapted to extract nucleic acids from all different types of microorganisms and matrices. 

T.-H. Wang, Y. Zhang, Fabrication of hierarchical silica nanomembranes and uses thereof for solid phase extraction of nucleic acids, US 2015/0,037,802 Al, 2015. 

After extraction, the RNA obtained from mammalian ribosomes separates into two groups—a 28 S and an 18 S RNA. The 28 S is derived from the two-thirds portion, and the 18 S from the one-third portion of the ribosome. Whether or not each of these two RNA s constitutes a single polynucleotide chain is not certain, but if they do, the molecular weight of the 28 S would be about twice that of the 18 S (1.2 x 10 and 0.6 x 10, respectively). Similarly, extraction of nucleic acid from ribosomal segments of E. coli yields 23 S and 16 S RNA base composition studies show significant differences between the two types of RNA. 

Elaissari A, Holt L, Meunier F, Voisset C, Pichot C, Mandrand B, Mabilat C. Hydrophilic and cationic latex particles for the specific extraction of nucleic acids. J. Biomater. Sci. Polym. Edn. 1999 10 403 20. 

Body fluids, extraction of nucleic acids from, 958... 

Measuring sensor, 378-379 Medical fluids, extraction of nucleic acids from, 958 Medical studies, application of TLC of indoles in, 912... 

The specific extraction of nucleic acid molecules is the only method of molecular biology-based diagnostic. Jiang et al. have reported the isolation of messenger ribonucleic acid (mRNA)... 

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