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194 producción de energía en una central termoeléctrica, o para refrigeración entre otros. diseñar sistemas de Actualmente hay una problemática en el mundo entero y es el control de la contaminación que a diario producen millones de automotores, turbinas de los aviones, maquinaria industrial, de tal manera que mejorar la eficiencia de las máquinas térmicas sin contaminar el medio ambiente representa un constante reto para la ingeniería. Por otra parte, en la industria de los alimentos la refrigeración es un proceso muy importante, ya que las bajas temperaturas inhiben el crecimiento microbiano y las reacciones enzimáticas, causantes de la descomposición de la mayoría de productos alimenticios. Surgen entonces varias preguntas, por ejemplo: cómo producir frío?, qué elementos constituyen un sistema de refrigeración?, qué características debe poseer un material para que sea un buen refrigerante? Es necesario precisar que todo proceso de refrigeración implica que exista un sistema que retire calor de una región de baja temperatura y lo lleve a otra de mayor temperatura. Como Ud. debe conocer, del estudio de la unidad anterior, este proceso no se realiza en forma espontánea y para que ocurra se necesita que el sistema cumpla un ciclo donde se realice trabajo sobre él y como resultado se verifique la transferencia de calor en el sentido deseado. Al final de la unidad, se profundizará en el estudio de los ciclos de refrigeración y las características de las sustancias utilizadas como refrigerantes. Lección : La máquina de vapor. Ciclo de Rankine En la segunda unidad de nuestro curso vimos cómo la máquina de vapor diseñada por Thomas Newcomen y perfeccionada por James Watt, mostraba la relación que podría existir entre trabajo y calor; y sirvió como idea base para que Joule proyectara el experimento del equivalente mecánico del calor. Dos siglos después del invento de Watt, existen todavía numerosas máquinas que funcionan con el mismo principio termodinámico original y que nos muestran la utilidad y vigencia de la máquina de vapor en nuestra sociedad. El ciclo termodinámico de esta máquina fue estudiado por W.J. Rankine, un ingeniero escocés que en 859 publicó su libro Manual de la Máquina de Vapor y que le dio su nombre al ciclo termodinámico de la máquina de vapor. El fundamento mecánico de esta máquina consiste en aprovechar la energía que posee el vapor de agua para mover un pistón dentro de un cilindro y a la utilización de dos elementos tan baratos como son el agua y el carbón. La figura 88 nos muestra un esquema muy sencillo de la máquina de vapor. El agua se lleva mediante una bomba hasta la caldera, en donde la presión y la temperatura que alcance el vapor saturado dependerán de las condiciones de operación propias de la
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The adequate planning of sequencing experiments so as to avoid technical biases is as important as good experimental design, especially when the experiment involves a large number of samples that need to be processed in several batches. In this case, including controls, randomizing sample processing and smart management of sequencing runs are crucial to obtain error-free data (Fig. 1a; Box 2).
When a reference genome is available, RNA-seq analysis will normally involve the mapping of the reads onto the reference genome or transcriptome to infer which transcripts are expressed. Mapping solely to the reference transcriptome of a known species precludes the discovery of new, unannotated transcripts and focuses the analysis on quantification alone. By contrast, if the organism does not have a sequenced genome, then the analysis path is first to assemble reads into longer contigs and then to treat these contigs as the expressed transcriptome to which reads are mapped back again for quantification. In either case, read coverage can be used to quantify transcript expression level (Fig. 1b). A basic choice is whether transcript identification and quantification are done sequentially or simultaneously.
Read mapping and transcript identification strategies. Three basic strategies for regular RNA-seq analysis. a An annotated genome is available and reads are mapped to the genome with a gapped mapper. Next (novel) transcript discovery and quantification can proceed with or without an annotation file. Novel transcripts are then functionally annotated. b If no novel transcript discovery is needed, reads can be mapped to the reference transcriptome using an ungapped aligner. Transcript identification and quantification can occur simultaneously. c When no genome is available, reads need to be assembled first into contigs or transcripts. For quantification, reads are mapped back to the novel reference transcriptome and further analysis proceeds as in (b) followed by the functional annotation of the novel transcripts as in (a). Representative software that can be used at each analysis step are indicated in bold text. Abbreviations: GFF General Feature Format, GTF gene transfer format, RSEM RNA-Seq by Expectation Maximization
PacBio RNA-seq is the long-read approach with the most publications to date. The technology has proven useful for unraveling isoform diversity at complex loci [186], and for determining allele-specific expression from single reads [187]. Nevertheless, long-read sequencing has its own set of limitations, such as a still high error rate that limits de novo transcript identifications and forces the technology to leverage the reference genome [188]. Moreover, the relatively low throughput of SMRT cells hampers the quantification of transcript expression. These two limitations can be addressed by matching PacBio experiments with regular, short-read RNA-seq. The accurate and abundant Illumina reads can be used both to correct long-read sequencing errors and to quantify transcript levels [189]. Updates in PacBio chemistry are increasing sequencing lengths to produce reads with a sufficient number of passes over the cDNA molecule to autocorrect sequencing errors. This will eventually improve sequencing accuracy and allow for genome-free determination of isoform-resolved transcriptomes.
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