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Genomics > Enzymes And Restriction Mapping > Flashcards

Enzymes And Restriction Mapping Flashcards

1
Q

What are the uses of genetic engineering?

A

Transgenic organisms and recombinant proteins

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2
Q

Give some examples of transgenic organisms

A

Disease models and improved agricultural yields

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3
Q

Give some examples of recombinant proteins

A

Insulin, interferon and G-CSF

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4
Q

What does G-CSF do?

A

Promote formation of bone marrow in cancer patients

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5
Q

What do nucleases do?

A

Degrade nuclear acids by hydrolysing phosphodiesterase bonds

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6
Q

What degrades RNA?

A

Ribonuclease

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7
Q

What degrades DNA?

A

Deoxyribonuclease

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8
Q

What do restriction endonucleases do?

A

Limits the transfer of nucleic acids from infecting phages into bacteria

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9
Q

How do restriction endonucleases work?

A

Recognising a specific sequence and cutting it.

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10
Q

What is special about restriction endonuclease recognition sites?

A

4-8 bases in length and palindromic

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11
Q

What does DNA ligase do?

A

Joins DNA molecules from different sources to form recombinant DNA

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12
Q

What can SNPs do?

A

Create/ destroy restriction enzyme sites

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13
Q

What do you use the enzyme Dde1 to do?

A

Cleave when doing PCR for sickle cell anaemia testing

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14
Q

What is a restriction map?

A

A crude way of mapping an unknown molecule

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15
Q

What is a restriction map useful for?

A

Describing plasmids

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16
Q

How do you get a restriction map?

A

By performing gel electrophoresis

17
Q

What are the main uses of DNA polymerases?

A

PCR amplification, generation of probes and blunt- ending of DNA

18
Q

What are phosphatases used for?

A

Preventing cleaved plasmids from resealing

19
Q

How do phosphatases work?

A

Hydrolysing a phosphate group off its substrate

20
Q

How do polynucleotide kinases work?

A

Adds phosphate to the 5’ hydroxyl group of DNA or RNA

21
Q

What are polynucleotide kinases used for?

A

To sensitively label DNA so that it can be traced using radioactively or fluorescently labelled ATP

22
Q

What is a probe?

A

A fragment of single stranded DNA or RNA that is complementary to the gene of interest

23
Q

How long are probes generally?

A

20-1000 bases

24
Q

How does reverse transcriptase work?

A

RNA dependant DNA polymerase that is isolated from RNA containing retroviruses. Then synthesises a DNA molecule that is complementary to an mRNA template using dNTPs

25
Q

What are the three types of primers for reverse transcription?

A

Random primers, oligoprimers and gene specific primers

26
Q

What is good about using random primers for reverse transcription?

A

Complementary DNA up to 700 bases but will cover the length of all the RNA molecules

27
Q

What is good about using oligoprimers for reverse transcription?

A

They’re useful for cloning cDNAs and their libraries but some might not be full length

Genomics (16 decks)

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