Effectiveness of flameless catalytic infrared radiation against life stages of three stored-product insect species in stored wheat

A bench top flameless catalytic infrared emitter was evaluated in the laboratory to disinfest wheat containing different life stages (ages) of the lesser grain borer, Rhyzopertha dominica; rice weevil, Sitophilus oryzae; and red flour beetle, Tribolium castaneum. The emitter produces infrared in the 3 to 7 um range. A noncontact infrared thermometer obtained real-time grain temperatures during exposures of uninfested and infested wheat containing various life stages of the three insect species. The grain temperatures attained were influenced by wheat quantity, distance from the emitter, and exposure time, which in turn influenced effectiveness against various life stages of the three species. In general, higher grain temperatures were attained in 113.5 g of wheat as opposed to 227.0 g, at 8.0 cm from the emitter surface rather than at 12.7 cm, and during a 60-sec exposure compared to a 45-sec exposure. Logistic regression indicated the probability of death of various life stages of R.  dominica, S. oryzae, and T. castaneum was temperature-dependent. About 99 to 100% mortality of all life stages of the three species occurred when the mean wheat temperatures were in the range of 108 to 114°C. The promising results show flameless catalytic infrared technology to be a viable option for disinfestation of stored wheat, provided such high temperatures do not affect grain quality.Keywords: Infrared radiation, Stored-product insects, Non-chemical method, Efficacy assessmen

Enhanced Signals and Fast Nucleic Acid Hybridization By Microfluidic Chaotic Mixing

Order from chaos: Microfluidic chaotic mixing shows a significant improvement in the equilibration time of hybridization in DNA microarrays over conventional techniques. These results illustrate a new concept for performing kinetics studies in microfluidic devices

Enhanced Signals and Fast Nucleic Acid Hybridization By Microfluidic Chaotic Mixing

Order from chaos: Microfluidic chaotic mixing shows a significant improvement in the equilibration time of hybridization in DNA microarrays over conventional techniques. These results illustrate a new concept for performing kinetics studies in microfluidic devices

Genomic DNA as a General Cohybridization Standard for Ratiometric Microarrays

Feature variability on ratiometric microarrays is accommodated by simultaneous cohybridization of a labeled reference standard with a labeled experimental sample. An optimal reference standard would provide full and equal representation for all array features from a given genome so that it would function on any array, would represent all features with similar signal intensity, and would be highly reproducible—both technically and biologically—from preparation to preparation and laboratory to laboratory. A low cost and a good shelf life are also highly desirable. Finally, providing for straightforward recovery of RNA prevalence information and for integration of data across multiple, initially unrelated studies would be significant advances over current methods. For virtually all ratiometric array studies published to date the reference standard has been some kind of RNA sample assembled from a number of different cell lines, tissues, or experimental time points. These RNA references fall short of the desired universality, uniformity, and reproducibility criteria, which then affect data quality and integration across studies. Also, the various mixed RNA standards cannot be used to derive RNA prevalence information from an experimental sample. In contrast, genomic DNA is a natural choice to meet all the criteria, although it has not yet been widely exploited for eukaryotic array experiments. Principal stumbling blocks have been achieving high enough absolute signals for large mammalian and plant genomes and finding a way to stabilize labeled DNA so that it can be stored and used with ease. This chapter describes two genomic DNA‐labeling methods that make it possible to use genomic DNA as a universal microarray cohybridization standard. The indirect labeling method permits production of a large quantity of a stable genomic DNA standard that can then be quality tested and stored frozen. This optimizes experimental consistency and significantly improves ease of use. This chapter also shows that the genomic DNA reference standard can deliver RNA prevalence measurements from ratiometric array platforms

Genomic DNA as a cohybridization standard for mammalian microarray measurements

A persistent design problem for ratiometric microarray studies is selecting the ‘denominator’ RNA cohybridization standard. The ideal standard should be readily available, inexpensive, invariant over time and from laboratory to laboratory, and should represent all genes with a uniform signal. RNA references (both commercial ‘universal’ and experiment- specific types), fall short of these goals. We show here that mouse genomic DNA is a reliable microarray cohybridization standard which can meet these criteria. Genomic DNA was superior in universality of coverage (>98% of genes from a 16 000 feature mouse 70mer microarray) to the Stratagene Universal Mouse Reference RNA standard. Ratios for genes in very low abundance in the Stratagene standard were more unstable with the Stratagene standard than with genomic DNA. Genes with mid-range, and therefore presumably optimal RNA denominator values, showed comparable reproducibility with both standards. Inferred ratios made between two different experimental RNAs using a genomic DNA standard were found to correlate well with companion, directly measured ratios (Spearman correlation coefficient = 0.98). The advantage in array feature coverage of genomic DNA will likely increase as newer generation microarrays include genes which are expressed exclusively in minor tissue or developmental domains that are not represented in mixed tissue RNA standards

Genomic DNA as a General Cohybridization Standard for Ratiometric Microarrays

Feature variability on ratiometric microarrays is accommodated by simultaneous cohybridization of a labeled reference standard with a labeled experimental sample. An optimal reference standard would provide full and equal representation for all array features from a given genome so that it would function on any array, would represent all features with similar signal intensity, and would be highly reproducible—both technically and biologically—from preparation to preparation and laboratory to laboratory. A low cost and a good shelf life are also highly desirable. Finally, providing for straightforward recovery of RNA prevalence information and for integration of data across multiple, initially unrelated studies would be significant advances over current methods. For virtually all ratiometric array studies published to date the reference standard has been some kind of RNA sample assembled from a number of different cell lines, tissues, or experimental time points. These RNA references fall short of the desired universality, uniformity, and reproducibility criteria, which then affect data quality and integration across studies. Also, the various mixed RNA standards cannot be used to derive RNA prevalence information from an experimental sample. In contrast, genomic DNA is a natural choice to meet all the criteria, although it has not yet been widely exploited for eukaryotic array experiments. Principal stumbling blocks have been achieving high enough absolute signals for large mammalian and plant genomes and finding a way to stabilize labeled DNA so that it can be stored and used with ease. This chapter describes two genomic DNA‐labeling methods that make it possible to use genomic DNA as a universal microarray cohybridization standard. The indirect labeling method permits production of a large quantity of a stable genomic DNA standard that can then be quality tested and stored frozen. This optimizes experimental consistency and significantly improves ease of use. This chapter also shows that the genomic DNA reference standard can deliver RNA prevalence measurements from ratiometric array platforms