W7.4.2

Question 1

EIA results (Quantitative)

Answer 1

”- Interpreted in comparison to a standard curve (a serial dilution of a known, purified antigen) in order to precisely calculate the concentrations of antigen in various samples
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Question 2

EIA results (Qualitative)

Answer 2

”- Used to achieve a “yes” or “no” answer indicating whether a particular antigen is present in a sample, as compared to a blank well containing no antigen
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Question 3

EIA results (semiqualitative)

Answer 3

Used to compare the relative levels of antigen in assay samples, since the intensity of signal will vary directly with antigen concentration

Question 4

polymerase chain reaction info

Answer 4

”- A primer-mediated, temperature-dependent technique for the enzymatic amplification of a specific DNA/RNA sequence
- A typical reaction involves a total of 25-42 cycles:
* Denaturation (94oC for 15s to 1 min)
* Primer hybridization (52oC for 15s to 2 min)
* Extension from primers (72oC for 15s to 3 min)
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Question 5

polymerase chain reaction how it works

Answer 5

”- Denaturation involves the conversion of double- stranded DNA into single stranded DNA
- Primers will flank the two sides of the target sequence and serve to define the ends of the DNA that has been targeted for amplification
- Amplification of the target DNA depends on the action of the DNA polymerase
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Question 6

Molecular - Hybridization

Answer 6

“Nucleic acid hybridization: formation of a double- stranded molecule from complementary single- stranded molecules
Probe: marked (or labeled) single-stranded sequence of nucleic acid that is complementary to the nucleic acid sequence to be detected
Target: detected nucleic sequence
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Question 7

Types of Hybridizations

Answer 7

FISH, solid-support, solution-phase

Question 8

FISH

Answer 8

Fluorescence in situ hybridization (FISH) (detection of specific nucleic acid sequences within structurally intact cells or tissues)

Question 9

Solid-support

Answer 9

(target is secured to a membrane or a microtiter tray and probe is free

Question 10

Solution-phase

Answer 10

both target and probe move around freely

Question 11

Flow cytometry

Answer 11

”- Utilizes light to count and profile cells in a heterogeneous fluid mixture
Basic steps:
* Stained sample cells are passed through a narrow channel; one at a time
* Light is used to illuminate the cells in the channel
* Series of sensors detect the types of light that are scattered or emitted from the cells (different wavelengths)
* Data acquired by the sensors is compiled and integrated to build a comprehensive picture of the sample
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