Component 2.7 - Genetic Techniques Flashcards
Explain what gives individuals a unique DNA profile and how this is used to make a DNA profile in the UK
1) An individuals DNA profile is different to that of another’s
2) Exons code for DNA and are similar because we are the same species
3) Introns have sequences called Short Tandem Repeats (blocks of repeated nucleotides) and the number of repeats is different in each individual
4) A number of STRs will be used to make a DNA profile
What 2 processes does DNA sequencing rely on?
1) The polymerase chain reaction (to amplify small sections of DNA rapidly)
2) Gel electrophoresis
What happens in the first step of the polymerase chain reaction?
1) Target DNA heated to 95 degrees so it splits
What happens in the second step of the polymerase chain reaction?
2) Solution cooled to 55 degrees and primers (which are complementary to the start of the sequence) attach to complementary bases on the single strand of DNA
What happens in the third step of the polymerase chain reaction?
3) Solution heated to 70 degrees - taq polymerase catalyses the synthesis of a complementary strand by adding complementary nucleotides and forming sugar-phosphate backbone (extension)
What happens to the DNA before it goes in the thermocycler?
1) The DNA solution is dissolved in a buffer
2) It is mixed with taq polymerase
3) It is mixed with nucleotides and short single stranded pieces of DNA complementary to the start of the DNA strand
Limitations of the polymerase chain reaction
- Any DNA that enters the system can get amplified
- Taq polymerase cannot correct itself like DNA polymerase
What happens to the DNA before it undergoes gel electrophoresis?
- The DNA is extracted and cut to make DNA fragments using restriction endonucleases
- DNA are loaded in wells at one end of the agarose
How are DNA fragments separated by gel electrophoresis?
1) Voltage is applied across the gel
2) Phosphate groups in the DNA backbone are negatively charged and attracted to positive electrode
3) Shorter fragments travel further than larger fragments as it easier for them to travel through pores in the gel
Uses of DNA profiling?
Siblings - if they have similar banding they can prove to be biological siblings
Paternity - bands on the DNA profile of the alleged father line up with the DNA profile of the child where the mother does not
Monozygotic twins - have identical banding patten
Dizygotic twins - don’t have an identical banding pattern
What are the pros of DNA profiling?
- Can be used on samples too small for blood testing
- Reversed wrongful convictions
- Can use urine/mouth swabs instead of blood (non invasive)
What are the cons of DNA profiling?
- Some people think that a request for DNA is a violation of privacy
- Computer databases with DNA profiles could get hacked
- Limited access (e.g no health insurers)
- DNA profiling may produce wrongful convictions e.g performed incorrectly, people may not be trustworthy
How can fragment size be estimated?
By running a DNA ladder alongside as it contains fragments of known size