07 Dot Blot Assay Flashcards
(40 cards)
It is also known as enteric fever, is a communicable disease, found more likely in man and occurs due to systemic infection mainly by Salmonella typhi organisms, a gram-negative bacterium.
● Typhoid fever
Typhoid fever, also known as enteric fever, is a communicable disease, found more likely in man and occurs due to systemic infection mainly by _____ typhi organisms, a ____ bacterium
Salmonella
gram-negative
Environmental exposure to Salmonella typhi may be more frequent in ____, presumably due to sex-linked differences in hygiene practices and dining-out behavior. But, it predominantly affects the __________
males
children of school-age or younger.
These diseases are spread through sewage contamination of food or water and through person-to-person contact. People who are currently ill and people who have recovered but are still passing the bacteria in their feces can spread ________. Thus, it is through the _______
Salmonella Typhi
fecal–oral route.
It is an acute generalized infection of the reticuloendothelial system, intestinal lymphoid tissue, and the gallbladder.
Typhoid fever
It is a potentially fatal multisystemic illness that causes nearly 220,000 deaths annually and 22 million illnesses per year.
Typhoid fever
Isolation of Salmonella from ________ is the most reliable means of confirming an infection. Blood culture is regarded as the gold standard for diagnosis and carry 70-75% diagnostic yield in the first week of illness
blood, urine or stool
______can be more quickly analyzed using dot blots or slot blots.
DNA and RNA
DNA and RNA can be more quickly analyzed using dot blots or slot blots. These procedures are applied to:
(3)
- Expression
- Mutation
- Amplification/deletion analyses
Techniques in which target or probe sequences are immobilized on a solid matrix.
Blots
It follows a similar principle to Western or Southern blotting, except that the sample is not blotted from a gel. Instead, samples are dotted directly onto a membrane before being probed for detection.
Dot Blot Technique
multiple samples are immobilized in a geometric array on a nitrocellulose or nylon membrane
Dot Blot Principle
______ is a method for detecting biological samples like proteins or nucleic acids.
Dot blot
Samples are dotted directly on to a membrane before being probed for detection.
The target DNA or RNA is deposited ____ on the membrane by means of various devices, some with vacuum systems.
directly
A _____can be used for procedures testing only a few samples.
pipet
For _____, the target is deposited in a circle or dot.
dot blots
_____ are useful for multiple qualitative analyses where many targets are being compared, such as in mutational screening
Dot blots
Performed most efficiently in less complex samples, such as cloned plasmids, PCR products, or selected mRNA preparations
dot blot
t
It is important that the probe hybridization conditions be optimized because cross-hybridizations cannot be definitively distinguished from true target identification.
A _______ serves as the baseline for interpretation of this dot blot assay.
negative control
Applications of Dot Blot:
- Monitoring labeling efficiency
- Estimating protein concentration
- Comparing antibody performance
Dot Blot Analysis:
- ImageJ
- UN-SCAN-IT gel Analysis Software
_____ plugin used to measure microarray image stacks. Use the control panel that appears to define the ROI (circle or rectangle) grid.
ImageJ
