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Molecular Biology & Genetics > MCQ 2 > Flashcards

MCQ 2 Flashcards

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1
Q

Southern blotting

A

A method to detect single stranded DNA that has been transferred to nylon paper by using a probe that binds DNA

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2
Q

Northern blotting

A

Hybridisation technique where a DNA probe binds to an RNA target molecule

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3
Q

Steps of Southern blotting

A

Target DNA cut with restriction enzyme
Fragments separated by gel electrophoresis
dsDNA melted by soaking gel in alkaline denaturing solution
DNA fragments transferred to nylon membrane
Membrane dipped in solution of labelled DNA probes molecules

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4
Q

What’s an example of a alkaline denaturing solution?

A

Sodium hydroxide

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5
Q

How do the DNA probe molecules work in Southern blotting?

A

They only bind to DNA fragments with similar sequences
Makes area radioactive
A black spot will appear where the hybrid molecule is if a piece of photographic film is placed on top

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6
Q

What makes the primers needed for PCR?

A

Primase

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7
Q

Why is SYBRR green a useful reagent for real time PCR?

A

It flouresces only when bound to dsDNA

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8
Q

What are the mains things needed in a PCR reaction mixture?

A

DNA polymerase
Primers
Deoxyribosenucleoside triphosphates

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9
Q

What type of template is needed for RT-PCR

A

RNA template

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10
Q

What type of template is needed for inverse PCR?

A

A circular template

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11
Q

Which variant of PCR may be used to selectively amplify mRNAs from a sample of total cellular RNA?

A

Differential display PCR

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12
Q

What are the steps and temperatures of DNA in order?

A

Strands melt @ 90oC
Primers anneal @ 50-60oC
DNA synthesis @70oC

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13
Q

What is responsible for DNA synthesis in PCR?

A

Taq polymerase

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14
Q

Inverse PCR

A

Method for using PCR to amplify unknown sequences by circularising the template molecule

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15
Q

When do you use inverse PCR?

A

When you know a bit of a DNA sequence in the middle but not the bits either side

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16
Q

Degenerate primer

A

A primer with several alternative bases at certain positions

17
Q

Long PCR

A

PCR reaction used specifically to amplify longer target sequences than standard PCR

18
Q

Hot start PCR

A

Taq polymerase is kept inactive at low temperatures by antibodies or blocking proteins to make sure template DNA is fully denatured

19
Q

Unique steps of inverse PCR

A

Restriction enzymes cut DNA sequence, but not known area
Fragment ligated so it circularises
Primers used that face outwards from known sequence
PCR run on circular DNA

20
Q

What does inverse PCR result in?

A

Strands of DNA with known sequence at the ends

21
Q

Randomly amplified polymorphic DNA (RAPD)

A

Method for testing genetic relatedness using PCR to amplify arbitrarily chosen sequences

22
Q

Reverse transcriptase PCR

A

Allows genes to be amplified and cloned intron free

Uses mRNA and reverse transcriptase to make copies (cDNA)

23
Q

Differential display PCR

A

Variant of RT-PCR that specifically amplifies mRNA from eukaryotic cells using oligo(dT) primers

24
Q

Oligo(dT) primer

A

Primer used on 3’ end of cDNA in differential display PCR

25
Q

Rapid amplification of cDNA ends (RACE)

A

RT-PCR based technique
Uses internal primers to speed to up copying
Can work 3’ to 5’

26
Q

Real time fluorescent PCR

A

Uses DNA binding probes that fluoresce more or only when bound to dsDNA
As more dsDNA is produced, fluorescence increases as more probe binds

27
Q

Multiplex PCR

A

Real time PCR reaction
Uses multiple fluorescent dyes on different probes
Assesses amplification of more than one target sequences

28
Q

Long interspersed element

A

Makes up majority of moderately repetitive DNA
Long sequences of DNA
Found in multiple copies

29
Q

Short interspersed element

A

Short repetitive sequences of DNA that are highly repetitive

30
Q

Satellite DNA

A

Highly repetitive DNA of eukaryotic cells
Found as long clusters of tandem repeats
Permanently coiled tightly into heterochromatin

31
Q

Tandem repeats

A

Repeated sequences of DNA/RNA that lie next to each other

32
Q

How many base pairs in a microsatellite?

A

~13

33
Q

How mant base pairs in a minsatellite?

A

~25

34
Q

Variable number tandem repeats (VNTR)

A

Cluster of tandemly-repeated sequences in DNA

Numbers of repeats differs from one individual to another

35
Q

What is used in DNA fingerprinting?

A

Variable number tandem repeats (VNTR)

Molecular Biology & Genetics (8 decks)

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