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A couple of PRNA 12-mers with desired purine pyrimidine mixed sequences were prepared indeed in high yields by the solid-phase synthesis.
Multiple infections revealed by mixed sequences were resolved by TA-cloning [27].
Mixed sequences were resolved by cloning the amplicon using the TOPO TA Cloning Kit (Life Technologies).
Mixed sequences were obtained with the isolate Poker Cat with spacers Cox5, Cox56, and Cox61.
These mixed sequences were closely related and were resolved by denaturing gradient gel electrophoresis (DGGE) giving characteristic band patterns for each species examined.
When mixed sequences were obtained, PCR amplicons were cloned using TOPO TA Cloning kits with the pCR 2.1-TOPO vector (Invitrogen, Carlsbad, CA) and sequencing was repeated using several clones.
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In the second stage, seven risk attributes against the shortlisted suppliers are further considered, and a new ranking method based on grey correlation degree of mixed sequence is proposed to rank the finalists and to select the final winners.
Such false mixed contiguous sequences were removed from the set of contiguous sequences.
The original flanking regions making up the false mixed contiguous sequences were then collected, and a contiguous sequence was rebuilt if there was more than one transcript-flanking region for a particular gene, whereas singlets were simply added to the final set.
The reaction products were gel purified and mixed, and contiguous sequences were generated by overlapping PCR using primers F1 and R2.
Previously, a series of de novo designed α-helical peptides with mixed Leu/Val sequences was presented, mimicking fusiogenically active transmembrane segments in model membranes (Hofmann et al., Proc. Natl. Acad. Sci. USA 101 (2004) 14776 14781).
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