MICRO EXAM 2 CH11

Question 1

What is nucleic acid hybridization?

Answer 1

The formation of hydrogen bonds between nucleotides of SS of RNA or DNA that are complementary to each other.

Question 2

What is duplex formation?

Answer 2

Two complementary strands of nucleic acids binding together to form a DS structure.

Question 3

What is the target in SS nucleic acid molecules?

Answer 3

Sequence that will be identified that is either immobilized or suspended in solution.

Question 4

What is the probe in SS nucleic acid molecules?

Answer 4

Short ssRNA or ssDNA oligonucleotide that is labeled with a chemical or fluorescent molecule; used to detect the target.

Question 5

What is probe selection?

Answer 5

Determines recognition to the target sequence and forms the duplex/hybrid with every complementary sequence.

Question 5

What is blotting?

Answer 5

Technique that transfer DNA or RNA or proteins from gel to membrane.

Question 5

Types of blots?

Answer 5

Northern, Southern and western blots.

Question 5

What is southern blot and its target?

Answer 5

DNA immobilized on solid support.
Uses restricition enzymes.

Question 6

What is northern blot and its target?

Answer 6

RNA (usually mRNA) immobilized on solid support.
Does not use restricition enzymes.

Question 7

How is protein extract separated in a western plot?

Answer 7

SDS-Page

Question 7

What is western plot and its target?

Answer 7

Proteins immobilized on solid support.

Question 8

How is protein blotted onto membrane?

Answer 8

Elecetrophoresis

Question 9

What does In situ hybridization detect?

Answer 9

Detects DNA/RNA (viruses ini tissue) directly in cells and tissue with labeled probes encased in paraffin (wax).

Question 10

What does NAAT stand for?

Answer 10

Nucleic Acid Amplification Test

Question 11

What does NAATs detect?

Answer 11

Amplifies the target microorganisms nucleic acid in a sample in a short amount of time.

Question 12

What does PCR stand for?

Answer 12

Polymerase Chain Reaction

Question 13

1st step in basic PCR cycle?

Answer 13

Denatures dsDNA

Question 13

Types of PCR methods?

Answer 13

Real Time-PCR, Reverse Transcriptase-PCR, multiplex PCR, Nested PCR

Question 14

2nd step in basic PCR cycle?

Answer 14

Annealing of primer

Question 15

3rd step in basic PCR cycle?

Answer 15

Extension of primer

Question 16

What is needed for PCR?

Answer 16

Template DNA
Oligonucleotide primers
Thermostable DNA pol

Question 16

How do you prevent contamination?

Answer 16

Having separate areas for each step, use of UV for decontamination, PPE

Question 17

How do you analyze PCR products?

Answer 17

Gel electrophoresis

Question 17

What does RT-PCR detect?

Answer 17

PCR amplicons (copies of DNA) after each cycle that has a fluorescence tag

Question 18

What is the purpose of the melting curve?

Answer 18

Determine results of verify purity of PCR products when 50% has been hybridized.

Question 19

What does Reverse Transcriptase PCR detect?

Answer 19

Detects mRNA products.

Question 20

What enzyme is used to convert mRNA to cDNA?

Answer 20

RNA-dependent DNA pol

Question 21

What does multiplex PCR detect?

Answer 21

Simultaneously detects two or more targets in the same sample

Question 22

What does nested PCR detect?

Answer 22

Amplifies first target region then amplifies another target region from the first amplification.

Question 23

What does MALDI-TOF stand for?

Answer 23

Matrix-Assisted Laser Desorption-Ionization Time Of Flight

Question 24

What does MALDI-TOF method use to detect microorganism?

Answer 24

Mass spectrometry

Question 25

1st step of MALDI-TOF?

Answer 25

Small amount of colony is applied to metal target.

Question 26

2nd step of MALDI-TOF?

Answer 26

Energy absorbing chemical. matrix solution is added.

Question 27

3rd step of MALDI-TOF?

Answer 27

Metal target is loaded into the ionization chamber.

Question 28

4th step of MALDI-TOF?

Answer 28

Matrix lattice is pulsed with a UV laser.