This report describes the development of a novel dual color Southern protocol to visualize two distinct genomes or genic regions simultaneously on a single Southern blot. The blot is developed with IRDye-conjugated antibody (Ab) and streptavidin that recognize digoxigenin (Dig)- or biotin-labeled probes, respectively and visualized on an infrared imager. This protocol was validated by visualizing viral and host genomes of human cytomegalovirus (HCMV)-infected human fibroblasts. This technique utilizes extremely sensitive fluorescent imaging, allowing the detection of nanogram quantities of DNA, as opposed to microgram quantities needed in Southerns using radioactively labeled probes, and eliminates the inherent loss in signal after stripping and reprobing a Southern blot. The probes are labeled with non-radioactive Dig and biotin and can be stored for extended periods of time. This protocol will aid in studies of any system with two genomes, such as cells infected with numerous types of microorganisms (virus/parasites/bacteria), or studies of mitochondrial and nuclear DNA within the same cells.
Pharmacology, Toxicology and Pharmaceutical Science
5
11%
Medicine and Dentistry
4
9%
Engineering
3
7%
Other
6
13%
Unknown
5
11%
Attention Score in Context
Attention Score in Context
This research output has an Altmetric Attention Score of 1. This is our high-level measure of the quality and quantity of online attention that it has received. This Attention Score, as well as the ranking and number of research outputs shown below, was calculated when the research output was last mentioned on 19 February 2014.
All research outputs
#23,677,354
of 28,879,505 outputs
Outputs from Journal of Virological Methods
#3,057
of 3,508 outputs
Outputs of similar age
#260,119
of 332,808 outputs
Outputs of similar age from Journal of Virological Methods
#15
of 17 outputs
Altmetric has tracked 28,879,505 research outputs across all sources so far. This one is in the 9th percentile – i.e., 9% of other outputs scored the same or lower than it.
So far Altmetric has tracked 3,508 research outputs from this source. They receive a mean Attention Score of 4.7. This one is in the 7th percentile – i.e., 7% of its peers scored the same or lower than it.
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We're also able to compare this research output to 17 others from the same source and published within six weeks on either side of this one. This one is in the 5th percentile – i.e., 5% of its contemporaries scored the same or lower than it.
