![]() 20, 21 Moreover, by affecting the kinetics of target/probe hybridization, surface oligonucleotide density plays an important role in the efficiency of duplex formation, as thermodynamic equilibrium may in some cases require excessively long incubation times. 16 – 19 The thermodynamic stability of double-stranded DNA is not just a function of the nucleic acid sequences, but is also affected by nearest-neighbor interactions or molecular crowding as determined in part by oligonucleotide density on the surface. The hybridization of complementary strands of DNA, which is the basis of all array-based techniques for nucleic acid analysis, is strongly dependent on surface oligonucleotide density both thermodynamically and kinetically. Surface oligonucleotide density is crucial for a wide variety of applications of DNA arrays. Sequence-recognition properties of DNA-binding proteins and small molecules have been determined on DNA arrays in a high-throughput fashion. Recent years have also seen emergence of the application of DNA arrays to molecular recognition studies. 10, 11 These studies are based on either hybridization of labeled nucleic acids to arrays or enzymatic reactions on array surfaces. Planar arrays of oligonucleotide probes have brought fundamental changes in nucleic acid analysis, including genotyping, 1, 2 resequencing, 3, 4 genome-scale expression analysis, 5, 6 single nucleotide polymorphism (SNP) detection, 7 – 9 and sequence capture and enrichment. It is shown that the procedure is reasonably general, in that it is readily transferable to alternative substrate materials, yielding similar results. Modulation of the UV exposure permits a desired degree of deprotection of surface synthesis sites a subsequent capping reaction to inactivate the exposed sites leaves only a desired fraction of active sites remaining for synthesis, corresponding to a lower oligonucleotide density. In this report, we describe an approach for the control of oligonucleotide density in photolithographically fabricated DNA arrays, based upon a controlled UV light deprotection procedure. A key parameter in the performance of DNA arrays is the density of the surface-bound oligonucleotides, which strongly affects both thermodynamic and kinetic aspects of DNA hybridization. Important application areas include whole-genome gene expression studies, high throughput analyses of single nucleotide polymorphisms, and most recently, the determination of binding site specificities for transcription factors and other critical elements involved in gene regulation. I highly recommend Richard Smith’s involvement to any individual or organisation looking for a personable, well-informed and supportive approach to enhancing development and achieving excellence.Over the last two decades high-density DNA arrays have developed into a central technology for nucleic acid analyses. Much of the input that he has provided to myself and to Snowsports New Zealand (formerly Winter Performance Programme) still influences our coach development programme, our coaching team values and our culture. I continue to seek feedback and support from Richard after meeting him over 15 years ago. Richard’s ability to apply theory into practice, his wealth of high-performance knowledge, his caring and empathetic nature were of high value to me and have helped guide my performance and development. ![]() Richard Smith stands out as one of the best, providing me with just the right level of support and challenge in my formative years as a high-performance coach. ![]() ![]() “I have been lucky enough to have many great mentors in my coaching career. ![]() Head Coach Park & Pipe Snowsports NZ High Performance Programme ![]()
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