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1993,59(2):327–329.PubMedCrossRef 50. Sambrook J, Maniatis T, Fritsch EF: Molecular cloning: A Laboratory Manual. Cold Spring Harbor, Cold Spring Harbor www.selleckchem.com/products/pu-h71.html Press; 1989. 51. Gradia DF, Rau K, Umaki AC, de Souza FS, Probst CM, Correa A, Holetz FB, Avila AR, Krieger MA, Goldenberg S, et al.: Characterization of a novel Obg-like ATPase in the protozoan Trypanosoma cruzi. Int J Parasitol 2009,39(1):49–58.PubMedCrossRef Authors’ contributions MB and FKM participated in the design of the platform, the cloning process, the validation of vectors and drafted the manuscript. PAFC carried out the TAP procedures and see more helped to draft the manuscript. SPF participated in the cloning process and the Southern blot analysis and contributed to scientific discussion. CMP participated in the DNA sequencing analysis, the cloning
process and contributed to scientific discussion. HP formatted the figures and contributed to vector validation. LSO, GAB and SG contributed to the design of the platform. MAK conceived the study, participated in the platform design and coordinated the project. All authors read and approved the final manuscript.”
“Background Microbial diversity in sediment or soil environments is very high, but the exact number of the taxa richness remains elusive . The estimated bacterial species ranged from nearly 103  to over 106  in a gram of sediment sample. Nevertheless, the figure has never been verified because of the low throughput of the traditional 16 S rRNA clone library method. Determining 16 S rRNA short variable tags using the pyrosequencing provided an unprecedented sequencing depth with tens to hundreds of thousands of tags per sample [4, 5], and the method regenerated FK866 cell line people’s interest in measuring and comparing the microbial taxa richness in various samples [6–8]. Nevertheless, two major types of problems about the 16 S rRNA pyrosequencing process were shortly revealed.