Underlying Data for Sequencing the Mitochondrial Genome with the Massively Parallel Sequencing Platform Ion Torrent (TM) PGM (TM)

Background: Massively parallel sequencing (MPS) technologies have the capacity to sequence targeted regions or whole genomes of multiple nucleic acid samples with high coverage by sequencing millions of DNA fragments simultaneously. Compared with Sanger sequencing, MPS also can reduce labor and cost on a per nucleotide basis and indeed on a per sample basis. In this study, whole genomes of human mitochondria (mtGenome) were sequenced on the Personal Genome Machine (PGM (TM)) (Life Technologies, San Francisco, CA), the out data were assessed, and the results were compared with data previously generated on the MiSeq (TM) (Illumina, San Diego, CA). The objectives of this paper were to determine the feasibility, accuracy, and reliability of sequence data obtained from the PGM. Results: 24 samples were multiplexed (in groups of six) and sequenced on the at least 10 megabase throughput 314 chip. The depth of coverage pattern was similar among all 24 samples; however the coverage across the genome varied. For strand bias, the average ratio of coverage between the forward and reverse strands at each nucleotide position indicated that two-thirds of the positions of the genome had ratios that were greater than 0.5. A few sites had more extreme strand bias. Another observation was that 156 positions had a false deletion rate greater than 0.15 in one or more individuals. There were 31-98 (SNP) mtGenome variants observed per sample for the 24 samples analyzed. The total 1237 (SNP) variants were concordant between the results from the PGM and MiSeq. The quality scores for haplogroup assignment for all 24 samples ranged between 88.8%-100%. Conclusions: In this study, mtDNA sequence data generated from the PGM were analyzed and the output evaluated. Depth of coverage variation and strand bias were identified but generally were infrequent and did not impact reliability of variant calls. Multiplexing of samples was demonstrated which can improve throughput and reduce cost per sample analyzed. Overall, the results of this study, based on orthogonal concordance testing and phylogenetic scrutiny, supported that whole mtGenome sequence data with high accuracy can be obtained using the PGM platform.Peer reviewe

Environmental and Genetic Preconditioning for Long-Term Anoxia Responses Requires AMPK in Caenorhabditis elegans

Article on environmental and genetic preconditioning for long-term anoxia responses requires AMPK in Caenorhabditis elegans

Developmental validation of the ForenSeq MainstAY kit, MiSeq FGx sequencing system and ForenSeq Universal Analysis Software

For human identification purposes, forensic genetics has primarily relied upon a core set of autosomal (and to a lesser extent Y chromosome) short tandem repeat (STR) markers that are enriched by amplification using the polymerase chain reaction (PCR) that are subsequently separated and detected using capillary electrophoresis (CE). While STR typing conducted in this manner is well-developed and robust, advances in molecular biology that have occurred over the last 15 years, in particular massively parallel sequencing (MPS) [1-7], offer certain advantages as compared to CE-based typing. First and foremost is the high throughput capacity of MPS. Current bench top high throughput sequencers enable larger batteries of markers to be multiplexed and multiple samples to be sequenced simultaneously (e.g., millions to billions of nucleotides can be sequenced in one run). Second, compared to the length-based CE approach, sequencing STRs increases discrimination power, enhances sensitivity of detection, reduces noise due to instrumentation, and improves mixture interpretation [4,8-23]. Third, since detection of STRs is based on sequence and not fluorescence, amplicons can be designed that are shorter in length and of similar lengths among loci, where possible, which can improve amplification efficiency and analysis of degraded samples. Lastly, MPS offers a single format approach that can be applied to analysis of a wide variety of genetic markers of forensic interest (e.g., STRs, mitochondrial DNA, single nucleotide polymorphisms, insertion/deletions). These features make MPS a desirable technology for casework [14,15,24, 25-48]. The developmental validation of the ForenSeq MainstAY library preparation kit with the MiSeq FGx Sequencing System and ForenSeq Universal Software is reported here to assist with validation of this MPS system for casework [49]. The results show that the system is sensitive, accurate and precise, specific, and performs well with mixtures and mock case-type samples.Peer reviewe

Surpat' chez Lolita / Les Orchestres : Jo Moutet ; Les Maimbas du Guatemala ; Manuel Pizarro ; Richard Audrey ; Eduardo Ruo

Titre uniforme : [Ciao, ciao, bambina = Piove]Titre uniforme : [Come softly to me]Titre uniforme : [Personality]Titre uniforme : [Carina]Titre uniforme : [Ciao, ciao, bambina = Piove]Titre uniforme : [Personnality]Titre uniforme : [Tout doux, tout doucement = Come softly to me]Comprend : Tout doux, tout doucement : fox / P. Delanoë ; F. Gérald ; B. Ellis ; Troxez ; Christopher - PERSONALITY : fox / Logan ; Price - LAS SECRETARIAS : cha cha cha / P. Luis - UNA CHICA CON IMPACTO : cha cha cha / F. Z. Maldonado - A MEDIA LUZ : tango / Donato - TANGO CUBANO / R. Calzado - Jazz tango / T. Fuggi ; Y. Allain - LUNA CAPRESE : boléro / L. Ricciardi - LOVE IN PORTOFINO : cha cha cha / J. Larue ; Chiosso ; Modugno - CIAO CIAO BAMBINA : cha cha cha / Verde ; Modugno ; Larue - NOCHE DE REYES : tango / Maffia - The gallant tango / J. Heyne - Cabaretera : boléro / B. Capo - UNA AVENTURA MAS : boléro / O. Kinleiner - TRUDIE : fox / J. Anderson - Carina : fox / PoesBnF-Partenariats, Collection sonore - BelieveContient une table des matière