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Biology-21-Recombinant DNA technology > Genetic Fingerprinting > Flashcards

Flashcards in Genetic Fingerprinting Deck (13)
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1

What is genetic fingerprinting

Is a diagnostic tool used widely in forensic science, plant and animal breeding, and medical diagnosis. It is based on the fact that the dna of every individual, expect identical twins, is unique

2

What are DNA bases that are non coding for

Variable number tandem repeats VNTRS

3

Can VNTRs be the same

They are different in all individuals except identical twins, and the probability of two individuals having identical sequences of these VTNRs is extremely small.

4

What is gel electrophoresis used to do

Separate DNA fragments according to their size

5

Describe how electrophoresis works

The DNA fragments are placed on to the afar gel and a voltage is applied across it.

The resistance of the gel means that the larger the fragments, the more slowly they move.

Therefore over a fixed period, the smaller fragments move further than the larger ones.

In this way DNA fragments of different lengths are separated

If DNA fragments are labelled for example with radioactive DNA probes, their final positions in the gel can be determined by placing a sheet of x ray Film over the ages gel for several hours.

The radioactivity from each dna fragment exposes the film and shows where the fragment is situated on the gel.

Only DNA fragments up to around 500 bases can be sequenced jn this way

Larger genes and whole genomes must therefore be cut into smaller fragments by restriction endonucleases

6

Describe the genetic fingerprinting technique

Extraction - dna is extracted from the sample (as amount of DNA is usually small, it’s quantity can be increased by using the polymerase chain reaction

Digestion - restriction endonucleases cut the DNA into fragments

Seperation - fragments are separated according to size by gel electrophoresis under the influence of an electrical voltage. DNA fragments are transferred from the gel to nylon membrane

Hybridisation- radioactive probes are now used to bind with VNTRs. The probes have base sequences which are complementary to the base sequences of the VNTRs, and bind to them under specific conditions such as temp and pH. The process is carried out with different probes, which bind to different target DNA sequences

Development - membrane with radioactively labelled dna fragments is placed onto an x ray Film. Development of the x ray Film reversals dark bands where the radioactive DNA probes have attached

7

How do we interpret the results from DNA fingerprints for example from blood at the scene of a crime and from a suspect

The pattern of bars of each fingerprint is passed through an automated scanning machine, which calculates the length of the DNA fragments from the bands.

It does this using data obtained by measuring the distances travelled during electrophoresis by known lengths of dna

Finally the odds are calculated of someone else having an identical fingerprint.

The closer the match between the two patterns, the greater the probability that the two sets of DNA have come from the same person

8

How can DNA fingerprinting be hard in resolving questions of paternity

As you inherit half of genetic material from Mum and half from dad.

Therefore each band on a DNA fingerprint of an individual should have a corresponding band in one of the parents

This can establish whether someone is the genetic father of a child

9

Even if there is a close match between a suspects DNA and the DNA found at a crime. What are the other possible explanations that need to be investigated

The DNA may have been left on some other, innocent occasion

The DNA may belong to a very close relative

The DNA sample may have been contaminated after the crime, either by the suspects DNA or by chemicals that affected the action of the restriction endonucleases used in preparing the fingerprint

10

Suggest how the genetic fingerprint of someone with the allele for Huntington’s disease might differ from that of someone who does not have the allele

In a person with the allele for Huntington’s disease, some of the dna fragments will be larger than those in a person without the allele because of the edges repeating units on the gene. These will travel a shorter distance in the electrophoresis gel and so there will be more thick bands nearest the start of the fingerprinting ( where the initial sample was located)


11

Explain how genetic fingerprinting can be used to ensure that interbreeding is avoided

Genetic fingerprinting can determine how closely any two individuals are related. The closer the match between their fingerprints, the close they are related. Therefore, to avoid the problems caused by interbreeding, it is advisable to mate animals whose fingerprints differ the most

12

Suggest how chemicals that affect the action of restriction endonucleases can alter the genetic fingerprint of a DNA sample

The chemicals may inhibit some of the restriction endonucleases, which would then fail to cut some sections of DNA. There would therefore be a greater number of longer DNA fragments than normal and the fingerprint would be different

13

Explain why it is often necessary to use the polymerase chain reaction when producing a genetic fingerprint

PCR is used to increase the quantity of DNA because the quantity available is often very small