Techniques in Molecular Biology

Question 1

The accumulation of amplification products is measured as the reaction progresses, in real time, with product quantification after each cycle.

Answer 1

Quantitative PCR (Qpcr)

Question 2

enabled by the inclusion of a fluorescent reporter molecule in each reaction well that yields increased fluorescence with an increasing amount of product DNA.

Answer 2

Quantitative PCR (Qpcr)

Question 3

Analytical tool in evaluating DNA copy number, viral load, SNP detection, and allelic discrimination

Answer 3

Quantitative PCR (Qpcr)

Question 4

Process in which DNA is cut at specific sites (restriction site), dictated by the surrounding DNA sequence.

Answer 4

Restriction Enzyme Digestion

Question 5

Used in molecular cloning techniques and PCR and AGE usually performed after

Answer 5

Restriction Enzyme Digestion

Question 6

Enzyme-linked immunosorbent assay

Answer 6

ELISA

Question 7

Labeled immunoassay and considered the gold standard of immunoassays

Answer 7

ELISA

Question 8

Performed in polystyrene plates, typically in 96-well plates coated to bind protein very strongly

Answer 8

ELISA

Question 9

Used to detect and quantify substances (antibodies, antigens, proteins, glycoproteins, and hormones).

Accomplished by complexing antibodies and antigens to produce a measurable result.

Answer 9

ELISA

Question 10

Testing requires a primary and/or secondary detection antibody, analyte/antigen, coating antibody/antigen, buffer, wash, and substrate/chromogen

Answer 10

ELISA

Question 11

4 main general steps in ELISA

Answer 11

1) Coating (either antigen or antibody),
2) blocking (typically by adding bovine serum albumin or BSA),
3) detection (adding substrate)
4) final read

Question 12

Target molecule of Southern blot

Answer 12

DNA

Question 13

Technique to detect a gene of interest in the DNA sample

Answer 13

Southern Blotting

Question 14

Used to detect proteins (including their size and amount of expression)

Answer 14

Western Blotting

Question 15

Visualization is made possible by primary and secondary antibodies

Answer 15

Western Blotting

Question 16

Used to detect specific RNA molecules, where after separation by electrophoresis, cDNA is used as a probe that binds to the RNA strand

Answer 16

Northern Blotting

Question 17

Utilize to measure RNA expression and size

Application includes finding alternate transcript size

Answer 17

Northern Blotting

Question 18

Extracted DNA or RNA or proteins is spotted onto the filter without the prior electrophoresis and transfer steps

Answer 18

Dot Blot

Question 19

Usually used for detection and quantification; Can also determines if an antibody is non-specific

Answer 19

Dot Blot

Question 20

Similar to the other blotting techniques, except it does not provide information regarding the size of the hybridized fragment

Answer 20

Dot Blot

Question 21

is the hybridization of fragmented single stranded DNA to arrays containing hundreds of thousands of unique nucleotide probe sequences.

Answer 21

SNP Microarray

Question 22

used to detect polymorphisms within a population (e.g., genetic abnormalities, breeding studies, etc.)

Answer 22

SNP Microarray

Question 23

Each probe is designed to bind to a target DNA subsequence.

Principle: DNA hybridization, fluorescence microscopy, and solid surface DNA capture

Answer 23

SNP Microarray