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Download Antisense and Ribozyme Methodology: Laboratory Companion by Ian Gibson PDF

By Ian Gibson

Antisense and ribozymes have a comparatively brief but winning heritage as study instruments in gene expression reports, and therefore are regarded as having excessive power reagents in treating viral infections and melanoma. This laboratory significant other offers specific details at the strength, benefits and barriers of this technique. It significantly discusses capability pitfalls, provides ideas for selecting ambitions and supply structures, with a purpose to enable the choice of the optimal method for reaching quickly and trustworthy experimental good fortune with any human or different organic process. For researchers, technicians and complicated graduates in experimental medication, molecular and telephone biology.

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Technically, microinjection is easiest with frog oocytes. However, in many cases mammalian tissue culture cells or primary cells are a more relevant biological system. Microinjection either into the nucleus or into the cytoplasm can be performed. Beside the advantages of microinjection, reproducibility and error range are sometimes unsatisfactory, which seems to be not only due to the cell culture conditions but also to the complexity of this technique. A standard deviation of microinjection experiments in the range of 15 % to 30 % is probably the best one can reach.

7 Endogenous Expression of Ribozyme Genes References Bratty J, Chartrand P, Ferbeyre G et a/. (1993):The hammerhead RNA domain, a model ribozyme. Biochim. Biophys. Acta 1216: 345-59 Cech TR (1993): Structure and mechanism of the large catalytic RNAs: Group I and group II introns and ribonuclease F! In The RNA World (Gesteland RF, Atkins JF, eds) pp 239-269, Cold Spring Harbor: Cold Spring Harbor Laboratory Press Crisell P, Thompson S, James W (1993):Inhibition of HIV-1 replication by ribozymes that show poor activity in vitro.

1986):Autolytic processing of dimeric plant virus satellite RNA. Science 231: 1577-80 Rhodes A, James WJ (1990) Inhibition of human immunodeficiency virus replication by endogenously synthesized antisense RNA. J. Gen. Virol. 71: 1965-74 The Design and Synthesis of Hammerhead Ribozymes Rittner K, Burmester C, Sczakiel G (1993):In vitro selection of fast-hybridizing and effective antisense RNAs directed against the human immunodeficiency virus type 1. Nucl. Acids Res. 21 : 1381-7 Sczakiel G, Homann M, Rittner K (1993): Computer-aided search for antisense RNA target sequences of the human immunodeficiency virus type 1.

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