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This protocol describes the TruSeq stranded mRNA preparation and sequencing for RNA-Seq experiments. The protocol is primarily focused on the generation of a stranded mRNA-Seq library from total RNA or polyA RNA extracts. While the steps are similar, data generated from both RNA fractions produce unique quality control metrics that need to be considered. This protocol is based on the two-step protocol developed by OATG (Illumina) for the preparation of TruSeq stranded mRNA libraries from total RNA extracts. The protocol also contains some variations, such as the duplex repair and the Enrichment of 3′ poly(A) fraction, which can be adapted to adapt the protocol to other mRNA-Seq kits.Purification and characterization of recombinant human lymphotoxin-α and its use in a bioassay for tumors of epithelial origin.
In this paper we report the successful cloning of the human lymphotoxin-α (LT-α) gene. The largest open reading frame identified was 632 amino acids (aa) long with an N-terminal signal sequence and a transmembrane domain. The human LT-α contained six conserved cysteines typical for a transmembrane protein. The protein was expressed as an inclusion body in Escherichia coli and the resulting inclusion body solubilized and refolded. The refolded LT-α was purified by ammonium sulphate precipitation and cation exchange chrom
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Among various kinds of semiconductor devices, a semiconductor device using gallium nitride (GaN), which has a wide band gap of 3.4 eV, has the properties such as a large breakdown voltage, a high efficiency, and a high-speed high power operation, in addition to the advantages of, for example, a high thermal conductivity and a high electron saturation velocity, and hence its application range is wide. Thus, it is expected that an application to high-frequency, high-power, and high-temperature semiconductor devices will be realized.
A semiconductor device using GaN is disclosed in, for example, the Journal of Crystal Growth, 93 (1989) 734-738, the publication of Japanese Patent Laid-Open No. H05-210458, and the publication of Japanese Patent Laid-Open No. 2000-242461. In addition, a semiconductor device using GaN is disclosed in, for example, Japanese Journal of Applied Physics, 35 (1996) L1220-1222, and the publication of Japanese Patent Laid-Open No. H03-183072.
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