Molecular Techniques Flashcards

1
Q

What are restriction enzymes?

A

Recognise and Cleave DNA at specific sites - commonly 4, 6, 8 base pair palindromes

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2
Q

What is the function of restriction enzymes in bacteria?

A

Protect DNA- degrades foreign DNA

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3
Q

What does DNA gel electrophoresis do?

A

Separate DNA fragments in an electric field based on size

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4
Q

What is a plasmid vector?

A

A small circular dsDNA found in bacteria

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5
Q

What’s the function of PCR?

A

To amplify specific DNA fragments

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6
Q

What three things does PCR require?

A

Thermostable taq polymerase
Primers (in pairs - reverse and forward)
Nucleotides

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7
Q

What three processes happen continually in PCR?

A

Denaturation
Annealing
Polymerisation

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8
Q

What does protein gel electrophoresis separate proteins on a basis of?

A

Size
Charge
Shape

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9
Q

What does SDS-PAGE do?

A

Separates proteins on size. SDS detergent denatures protein (removes secondary and tertiary structure).

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10
Q

What does isoelectric focusing (IEF) do?

A

Separates proteins based on pI - proteins move through gradient of pH. Migrate unruly pH = pI (no net charge)

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11
Q

What is 2D PAGE?

A

Separates on charge and size. Separates the bands from IEF into individual proteins

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12
Q

What is proteomics?

A

Analysis of all proteins in the genome and their functions

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13
Q

What is western blotting?

A

Following protein gel electrophoresis, bands are transferred onto a membrane (nitrocellulose) and incubated with primary antibody and then labelled secondary antibody

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14
Q

What is southern hybridisation?

A

Identifying specific DNA sequences using allele specific labelled probes

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15
Q

What is northern hybridisation?

A

Same as southern but uses RNA on gel

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16
Q

What is reverse transcriptase PCR?

A

Starts with mRNA - uses reverse transcriptase to produce cDNA then use PCR to amplify dsDNA

17
Q

What is karyotyping?

A

Lining up of chromosomes based on specific banding pattern

18
Q

What is FISH?

A

Uses specific labelled DNA probes to hybridise and fluoresce a particular part on a chromosome