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DNA Analysis & Interpretation > How to generate DNA profiles > Flashcards

How to generate DNA profiles Flashcards

1
Q

What is RFLP?

A

Restriction fragment length polymorphisms
* first DNA testing used
* took about a month to generate profiles
* separated by size
* very discriminating
* used radiation to mark where the DNA bands are

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2
Q

What is DQ Alpha?

A
  • first PCR-based test
  • look for the absence of blue dots - would show what sort of DNA molecules are present
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3
Q

What are the benefits of DQ Alpha over RFLP?

A
  • faster
  • more sensitive
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4
Q

What is MLP?

A

Multi-Locus Probe
* lengthy
* requires a large amount of sample to produce a profile
* results were complex to interpret
* only good for comparison

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5
Q

What is SLP?

A

Single Locus Probes
* easier to interpret
* intial results obtained in a few days but took over a week to prepare a full profile
* lacked sensitivity
* poor results with degraded samples
* difficult to resolve mixtures

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6
Q

What is sensitivity?

A

The amount of starting material or size of the stain we can begin working with

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7
Q

What is discriminating power?

A

The power to distinguish between one thing and another thing

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8
Q

What is the six criteria used for judging the benefits of a DNA profiling system?

A
  1. distriminating power
  2. sensitivity
  3. ability to deal with artefacts
  4. speed
  5. ability to deal with mixtures
  6. ability to conduct database searches
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9
Q

What is mitochondrial DNA?

A
  • more sensitive than automated STRs
  • every cell within it contains two copies of all of our genetic material
  • relatively small compared to the amount of DNA that is present in the nucleus so not much information is contained within
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10
Q

What is another drawback of mitochondrial DNA?

A
  • it is maternally inherited
  • will be many more maternal relatives who will share the genetic material
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11
Q

What is Y-STR?

A
  • tests on Y STR markers which reside on the human Y chromosome
  • can give us insight into the male DNA profile without seeing contributers from the female
  • helpful when there is a mixture
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12
Q

How much DNA do we need for DNA17?

A

500pg

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13
Q

What are polymorphisms?

A
  • non-coding part of DNA
  • differ greatly from person to person
  • the parts that are in DNA profiles
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14
Q

Primers in the PCR process?

A
  • forward and reverse primers
  • attached to one primer is a fluorescent dye
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15
Q

What does taq polymerase do?

A
  • is thermally stable
  • synthesises new DNA strands by extending primers along the template DNA
  • binds to the template strand at the start of the area for amplification
  • doesnt proceed until the temperature is increased (optimised temperature)
  • extension phase where the DNA polymerase synethesises new DNA strand by extending the primers along the whole amplification area
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16
Q

What is the function of dNTPs?

A
  • building blocks for the new DNA strand
  • in the extension step, DNA polymerase adds free dNTPs from the reaction mixture to the template DNA
17
Q

What are STRs?

A

Short Tandem Repeats
* small pieces of the genetic code which are repeated

18
Q

What is the vertical axis and horizontal axis represent in the raw data produced from capillary electrophoresis?

A
  • Vertical axis = relative fluorescence units, measures of the intensity of light picked up by the camera in the detector window at different points in time
  • Horizontal axis = minutes or seconds
19
Q

What are the first peaks in the raw data from capillary electrophoresis?

A

primers left over from the PCR process

20
Q

Where are the allelic drop out and drop in seen in an EPG?

A

at the end because they are the bigger bits so take longer to travel through and are more susceptible to degradation

21
Q

What is on the left and right side of the EPG?

A
  • left side: small fragments
  • right ride: large fragments
22
Q

What can be determine from the peak height in EPGs?

A
  • proportional to the amount of DNA that gave rise to that particular peak
23
Q

During the process of purification, where are the impurities to be found after centrifugation?

A

Pellet

24
Q

What is heterzygous?

A

two different alleles at a locus

25
Q

What are STRs and what are they also known as?

A

Short Tandem Repeats - mini/micro-satellites
* small pieces of genetic code that are repeated

DNA Analysis & Interpretation (17 decks)

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