Advanced Molecular Techniques Flashcards

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1
Q

Name 3 types of gene level DNA sequencing

A
  • Southern Hybridisation
  • Northern Hybridisation
  • Microarray
  • RT-PCR
  • DNA fingerprinting
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2
Q

Which enzyme does RNA PCR use?

A

Reverse transcriptase

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3
Q

What primer sequence is used on cDNA in RT-PCR and why?

A
  • Primer has T-tail

- Complementary mRNA has polyA tail

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4
Q

What are the 3 main steps in RT-PCR?

A

mRNA > cDNA > RT-PCR

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5
Q

What is the main use of microarray technology?

A

To compare the gene expression of two conditions or individuals with/without conditions

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6
Q

What is microarray technology a measure of?

A

Conditional gene expression

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7
Q

What do colours infer in microarray technology?

A

Colour demonstrates the ratios of cDNA that has bound from each sample

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8
Q

What are the 2 main steps in microarray technology involving mRNA?

A
  • mRNA isolation
  • Reverse transcriptase labelling
  • RNA is copied and makes cDNA with a fluorescent tag
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9
Q

What type of microarray technology compares an affected genome with a healthy genome?

A

Array Comparative Genome Hybridisation

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10
Q

What does array comparative genome hybridisation highlight?

A
  • Microdeletions

- Microduplications

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11
Q

What does DNA fingerprinting examine?

A

The non-coding part of the genome

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12
Q

What does DNA fingerprinting compare?

A

The allelic variation at each locus

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13
Q

What differs at each loci between individuals?

A

The number of mini-satellites

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14
Q

What advanced molecular technique is used to form DNA databases?

A

DNA fingerprinting

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15
Q

What is karyotyping?

A

Looks at the banding patterns in a full set of chromosomes

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16
Q

What does the molecular technique FISH stand for?

A

Fluorescent In Situ Hybridisation

17
Q

What are the 3 main steps in FISH?

A
  • DNA probe labelled with fluorescent dye
  • DNA denatured
  • DNA hybridised with a complementary sequence
18
Q

What 2 genetic components can be examined using FISH?

A
  • DNA

- Chromosomes

19
Q

What kind of changes does FISH identify?

A

Small changes such as microdeletions