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Because miRNAs were not included in the total RNA population during RNA isolation, we did not observe multiple hybridization bands that may result from pri-miRNA processing.
We also noticed that one out of five offspring from F0 #38 showed multiple hybridization bands (about 1 and 2.6 kb) indicating DNA rearrangement (Fig. 7).
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Variability in the signal intensities of the multiple Southern hybridization bands raises the possibility that Euglena contains a naturally aneuploid chromosome complement.
A complex and reproducible pattern of multiple hybridizing bands is produced in Southern hybridization experiments with a Euglena U3 snoRNA gene probe (Fig. 1A), implying that this gene is present in multiple copies in the Euglena genome.
Lines 60 and 37 displayed multiple hybridization-positive bands (4 and 2 respectively), while lines 231 and 308 displayed a single positive band.
Positive hybridization bands were noted regardless of the DNA source.
Visualization of hybridization bands was carried out using X-ray film (Kodak).
For semi-quantifying the intensity of hybridization bands, the ImageJ.exe program (http://rsb.info.nih.gov) was used.
These probes were used for Southern blot analysis to search for abnormal, patient-specific hybridization bands.
One to three hybridization bands were detected for the DNA from various rice accessions digested with different restriction enzymes under high-stringency hybridization.
While multiple hybridization replicates provided ∼97% accuracy, even a single replicate provided ∼95% accuracy.
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