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.
We report photoluminescence (PL) results obtained on p-type ZnSe epilayers grown by molecular beam epitaxy. As an acceptor dopant, we used an active nitrogen beam produced by a free radical nitrogen source. On the basis of a detailed analysis of PL data we propose a simple semiquantitative method for a quick and contactless evaluation of the net acceptor concentration in p-type ZnSe. In particular, we show that the intensity ratio of the donor–acceptor pair (DAP) emission to the acceptor-bound exciton (I1) emission strongly depends on both the excitation power and the quality of the sample, and because of that it cannot by itself be regarded as a good measure of the net acceptor concentration. On the other hand, the intensity of the DAP emission under saturation excitation shows a simple direct proportionality to the net acceptor concentration, thus providing a reliable tool for determining the relative doping level in p-type ZnSe films.
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