PCR and 16S Gene Flashcards

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1
Q

What is the primary structure of DNA?

A

backbone and nitrogen-containing bases

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2
Q

What is the backbone made of?

A

sugar and phosphate groups of deoxyribonucleotides (bases)

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3
Q

Where do nitrogen-containing bases project from?

A

from back of backbone

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4
Q

Does DNA have directionality?

A

yes

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5
Q

What are the 2 ends of DNA named?

A

3’ and 5’

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6
Q

What are the characteristics of the 3’ end?

A

has exposed group on 3’ carbon deoxyribose

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7
Q

What are the characteristics of the 5’ end?

A

5’ carbon phosphate group

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8
Q

What does DNA synthesis occur after?

A

creation of a phosphodiester bond

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9
Q

What is polymerase chain reaction (PCR)?

A

technique used to amplify specific regions of DNA for sequencing and genetic analysis

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10
Q

What are the 3 steps of PCR?

A

Denaturation
annealing
elongation

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11
Q

What temperature is denaturation?

A

95 degrees celsius

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12
Q

What temperature is annealing?

A

68 degrees celsius

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13
Q

What temperature is elongation?

A

72 degrees celsius

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14
Q

Where do ribosomes coordinate protein synthesis?

A

from an RNA template

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15
Q

What are the 2 ribosome subunits?

A

prokaryotes and eukaryotes

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16
Q

What do prokaryotes have?

A

23S, 5S, 16S

17
Q

What do eukaryotes have?

A

28S, 5.8S, 5S, 18S

18
Q

Why do we use 16S sequence?

A

to identify microbes

19
Q

What are chromosomes?

A

large pieces of DNA with many genes

20
Q

What is denaturation?

A

heat briefly to separate DNA strands

21
Q

What is annealing?

A

cool to allow primers to bond hydrogen bond with ends of target sequence

22
Q

What is elongation?

A

DNA polymerase adds nucleotides to 3’ end of each primer

23
Q

What is the importance of the primers in a PCR reaction?

A

they act as the starting point for DNA synthesis
defines the region that will be amplified

24
Q

What is a primer?

A

short fragment of single-stranded DNA that binds to one strand of DNA template and is recognized by DNA polymerase as the starting point to copy DNA

25
Q

What is agarose gel electrophoresis?

A

separates DNA fragments based on charge (positive or negative), size in base pairs (bp), and conformation (linear vs circular)

26
Q

What charge is DNA?

A

partial negative on the phosphate group of the backbone

27
Q

Where do DNA molecules loaded in gel migrate?

A

towards positive electrode

28
Q

How fast to smaller molecules travel?

A

rapidly

29
Q

What speed to larger molecules travel?

A

they are slowed down as they bump into gel matrix

30
Q

What is PCR product?

A

contains millions of copies of one specific stretch of DNA

31
Q

What is a DNA template?

A

any DNA molecule such as bacterial chromosome, plasmid, and fragment of DNA