DNA sequence information underpins genetic research, enabling discoveries of important biological or medical benefit. Sequencing projects have traditionally employed long (400–800 bp) reads, but the existence of reference sequences for the human and many other genomes makes it possible to develop new, fast approaches to re-sequencing, whereby shorter reads are compared to a reference to identify intra-species genetic variation. We report an approach that generates several billion bases of accurate nucleotide sequence per experiment at low cost. Single molecules of DNA are attached to a flat surface, amplified
in situ
and used as templates for synthetic sequencing with fluorescent reversible terminator deoxyribonucleotides. Images of the surface are analysed to generate high quality sequence. We demonstrate application of this approach to human genome sequencing on flow-sorted X chromosomes and then scale the approach to determine the genome sequence of a male Yoruba from Ibadan, Nigeria. We build an accurate consensus sequence from >30x average depth of paired 35-base reads. We characterise four million SNPs and four hundred thousand structural variants, many of which are previously unknown. Our approach is effective for accurate, rapid and economical whole genome re-sequencing and many other biomedical applications.
A theoretical model, based on the geometrical optics approach, has been developed to simulate various aspects of the phase Doppler particle analyzer (PDPA). The model has taken into consideration the nonuniform (Gaussian) illumination of the particles as they pass through the measurement probe volume. Instrument response curves have been generated for various scattering angles by performing spatial and temporal integration of the scattered intensity distribution over the receiver surface. Experimental and theoretical investigations have established the applicability of this instrument to both forward scattered and backscattered angles.
A theoretical model based on the Lorenz-Mie theory was used to study the response characteristics of the Aerometrics phase Doppler particle analyzer (PDPA). The validity of the model was verified experimentally, and its suitability for calculating measurement uncertainties was established. The theoretical and experimental results suggest that size resolutions of the order of +/-0.3 microm are possible when the PDPA is used to measure small spherical particles (< 10 microm). We show that the optical configuration of the PDPA plays an important role in establishing the sizing uncertainty of the instrument.
Planar laser scattering (PLS) and planar laser-induced fluorescence (PLIF) techniques are currently being used for rapid characterization of fuel sprays associated with gas turbine atomizers, diesel injectors, and automotive fuel injectors. These techniques can be used for qualitative, quantitative, and rapid measurement of fuel mass, spray geometry, and Sauter mean diameters in various sprays. The spatial distribution of the fuel mass can be inferred directly from the PLIF image, and the Sauter mean diameter can be measured by simultaneously recording the PLIF and PLS images and then ratioing the two. A spray characterization system incorporating the PLS and/or PLIF techniques has been loosely termed an optical patternator, and in this study, it has been used to characterize both steady and pulsed sprays. The results obtained with the optical patternator have been directly validated using a phase Doppler particle analyzer (PDPA).
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