probes for chromosomes 13, 18, and 21 were modified from the original probe-mix of the P095 kit (see Fig.
We used PCR amplification and Southern blot hybridization
to detect 9 virulence factors among 40 EHEC type-strains from the STEC Center, National Food Safety and Toxicology Center, Michigan State University (East Lansing, MI, USA).
An advanced feature of PerkinElmer's HybArray 12 is the option to increase the hybridization
area to 0.
9 (ECkruH678) (a) Signal intensity is the mean signal from separate hybridizations
performed on 2 different slides printed with 8 spots for each capture probe expressed as a percentage of the control oligonucleotide signal.
Thus, the results of the hybridization
conducted with mesocosm rRNA extracts support the idea that a Syntrophus-like population in the sediment culture was involved in 3-CB biodegradation, as suggested by the DGGE and sequencing analyses performed during the preliminary microcosm experiment.
The widely used Affymetrix technology holds the standardization advantage with respect to probes, hybridization
protocols, and data quantification (6).
The goal of this study was to demonstrate that transcript-dependent dye bias is consistent and measurable on a given day and can therefore be corrected mathematically using a pair of split-control hybridizations
(see "Terminology" in "Materials and Methods") to achieve a similar high level of technical accuracy.
Classic DNA microarray formats, such as Affymetrix's Genechip or custom microarrays on glass slides, require hybridization
times of ~18 h for detection of nucleic acids and are thus too slow for the rapid diagnosis of infectious diseases.
Finally, sequences must be examined for hybridization
characteristics, as abnormally high or low G + C content may skew the signal for that target relative to other targets.
To determine whether several hybridizations
of replicates performed better than hybridization
of pooled replicates, we performed two distinct reverse transcription reactions with our sample template and three independent RAP-PCR reactions for each reverse transcription, yielding six PCR probes (Fig.
We used albino frogs for in situ hybridization
studies of Xslug mRNA expression and pigmented frogs for all other studies.