Automated SNP genotype clustering algorithm to improve data completeness in high-throughput SNP genotyping datasets from custom arrays

High-throughput SNP genotyping platforms use automated genotype calling algorithms to assign genotypes. While these algorithms work efficiently for individual platforms, they are not compatible with other platforms, and have individual biases that result in missed genotype calls. Here we present data on the use of a second complementary SNP genotype clustering algorithm. The algorithm was originally designed for individual fluorescent SNP genotyping assays, and has been optimized to permit the clustering of large datasets generated from custom-designed Affymetrix SNP panels. In an analysis of data from a 3K array genotyped on 1,560 samples, the additional analysis increased the overall number of genotypes by over 45,000, significantly improving the completeness of the experimental data. This analysis suggests that the use of multiple genotype calling algorithms may be advisable in high-throughput SNP genotyping experiments. The software is written in Perl and is available from the corresponding author

Can models for forest attributes based on airborne laser scanning be generalized for different silvicultural management systems?

In Finland, interest in continuous cover forestry (CCF) has increased rapidly in recent years. During those years CCF has been examined from various viewpoints but not from the perspective of forest inventories. This holds especially true for applications based on remote sensing. Conversely, airborne laser scanning (ALS) data have been widely used to predict forest characteristics such as size distribution and vertical forest structure, which are closely related to the forest information needs of CCF. In this study we used the area-based approach to predict a set of stand attributes from ALS data (5 pulses per m2) in a CCF forest management experiment in Katajama & BULL;ki, eastern Finland. In addition to the CCF stands, the experiment included shelterwood stands and untreated stands. The predicted attributes included volume, biomass, basal area, number of stems, mean diameter, Lorey's height, dominant height, standing dead wood volume, parameters of the theoretical stem diameter distribution model, understory height and number of understory stems. Our main aim was to test whether the same model could be used across different management systems. The accuracy of the attributes predicted for the CCF stands was compared with the predictions for the other management systems in the same experiment. We also compared and discussed our results in relation to the even-aged stand attribute predictions that were conducted by using separate operational forest data collected from sites surrounding Katajama & BULL;ki. The results showed that forest data from the different management systems could be combined into a single model of a stand attribute, i.e., ALS metrics were found to be suitable for comparing different management systems in regard to differences in forest structure. The accuracy of the predicted attributes in the CCF plots was comparable to that of the other management alternatives in the experiment. The accuracy was also comparable to that of even-aged forests. The results of this study were promising; the stand attributes of CCF-managed forests could be predicted analogously to those of other management systems. This indicates that for the purposes of forest inventories there may not be a need to stratify forest lands by management system. It should be noted, however, that the study area was relatively small, that the forest stands were harvested in the 1980 s, and that the attributes may not have been completely exhaustive for CCF

Synchroneity of major late Neogene sea level fluctuations and paleoceanographically controlled changes as recorded by two carbonate platforms

Shallow-water carbonate systems are reliable recorders of sea level fluctuations and changes in ambient seawater conditions. Drilling results from Ocean Drilling Program (ODP) Legs 133 and 166 indicate that the timing of late Neogene sedimentary breaks triggered by sea level lowerings is synchronous in the sedimentary successions of the Queensland Plateau and the Great Bahama Bank. This synchrony indicates that these sea level changes were eustatic in origin. The carbonate platforms were also affected by contemporary, paleoceanographically controlled fluctuations in carbonate production. Paleoceanographic changes are recorded at 10.7, 3.6, and 1.7–2.0 Ma. At the Queensland Plateau, sea surface temperature shifts are documented by shifts from tropical to temperate carbonates (10.7 Ma) and vice versa (3.6 Ma); the modern tropical platform was established at 2.0–1.8 Ma. At Great Bahama Bank, changes were registered in compositional variations of platform-derived sediment, such as major occurrence of peloids (3.6 Ma) and higher rates of neritic carbonate input (1.7 Ma). The synchroneity of these changes attests to the far-field effects of modifications in the oceanographic circulation on shallow-water, low-latitude carbonate production

Development of a single tube 640-plex genotyping method for detection of nucleic acid variations on microarrays

Detection of DNA sequence variation is critical to biomedical applications, including disease genetic identification, diagnosis and treatment, drug discovery and forensic analysis. Here, we describe an arrayed primer extension-based genotyping method (APEX-2) that allows multiplex (640-plex) DNA amplification and detection of single nucleotide polymorphisms (SNPs) and mutations on microarrays via four-color single-base primer extension. The founding principle of APEX-2 multiplex PCR requires two oligonucleotides per SNP/mutation to generate amplicons containing the position of interest. The same oligonucleotides are then subsequently used as immobilized single-base extension primers on a microarray. The method described here is ideal for SNP or mutation detection analysis, molecular diagnostics and forensic analysis. This robust genetic test has minimal requirements: two primers, two spots on the microarray and a low cost four-color detection system for the targeted site; and provides an advantageous alternative to high-density platforms and low-density detection systems

Real-time DNA microarray analysis

We present a quantification method for affinity-based DNA microarrays which is based on the real-time measurements of hybridization kinetics. This method, i.e. real-time DNA microarrays, enhances the detection dynamic range of conventional systems by being impervious to probe saturation in the capturing spots, washing artifacts, microarray spot-to-spot variations, and other signal amplitude-affecting non-idealities. We demonstrate in both theory and practice that the time-constant of target capturing in microarrays, similar to all affinity-based biosensors, is inversely proportional to the concentration of the target analyte, which we subsequently use as the fundamental parameter to estimate the concentration of the analytes. Furthermore, to empirically validate the capabilities of this method in practical applications, we present a FRET-based assay which enables the real-time detection in gene expression DNA microarrays

A dual-tag microarray platform for high-performance nucleic acid and protein analyses

DNA microarrays serve to monitor a wide range of molecular events, but emerging applications like measurements of weakly expressed genes or of proteins and their interaction patterns will require enhanced performance to improve specificity of detection and dynamic range. To further extend the utility of DNA microarray-based approaches we present a high-performance tag microarray procedure that enables probe-based analysis of as little as 100 target cDNA molecules, and with a linear dynamic range close to 105. Furthermore, the protocol radically decreases the risk of cross-hybridization on microarrays compared to current approaches, and it also allows for quantification by single-molecule analysis and real-time on-chip monitoring of rolling-circle amplification. We provide proof of concept for microarray-based measurement of both mRNA molecules and of proteins, converted to tag DNA sequences by padlock and proximity probe ligation, respectively

A Rapid, Cost-Effective Method of Assembly and Purification of Synthetic DNA Probes >100 bp

Here we introduce a rapid, cost-effective method of generating molecular DNA probes in just under 15 minutes without the need for expensive, time-consuming gel-extraction steps. As an example, we enzymatically concatenated six variable strands (50 bp) with a common strand sequence (51 bp) in a single pool using Fast-Link DNA ligase to produce 101 bp targets (10 min). Unincorporated species were then filtered out by passing the crude reaction through a size-exclusion column (<5 min). We then compared full-length product yield of crude and purified samples using HPLC analysis; the results of which clearly show our method yields three-quarters that of the crude sample (50% higher than by gel-extraction). And while we substantially reduced the amount of unligated product with our filtration process, higher purity and yield, with an increase in number of stands per reaction (>12) could be achieved with further optimization. Moreover, for large-scale assays, we envision this method to be fully automated with the use of robotics such as the Biomek FX; here, potentially thousands of samples could be pooled, ligated and purified in either a 96, 384 or 1536-well platform in just minutes

Usability aspects of medication-related decision support systems in the outpatient setting: A systematic literature review

In this study, we evaluated the usability aspects of medication-related clinical decision support systems in the outpatient setting. Articles published