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DNA
In this IBDP Biology topic, you can learn different biotechnology method.
- Introns – the regions of the chromosomes which are used in DNA profiling.
- Mini-satellites – 20-50 base sequence repeated from 50 to several hundred times.
- Micro-satellites – 2-4 bases repeated between 5 and 15 times.
- The more closely related the two individuals, the more similar the DNA patterns are.
The process
- DNA is extracted from a blood or cell sample.
- It is split into fragments using restriction enzyme.
- These cut the DNA at certain points in the intron sequences.
- Each type cuts a DNA molecule into fragments at different recognition sites.
Gel electrophoresis
- The fragments are placed in wells in an agarose gel medium in a buffering solution, with known DNA fragments.
- The gel contains a dye which binds to the DNA fragments in the gel.
- A dye is also added to the DNA samples.
- An electric current is passed through the apparatus and the DNA fragments move towards the positive anode.
- The plate is placed under UV light.
- The DNA fluoresces so it can be identified.
Southern blotting
- An alkaline buffer solution is added to the gel after electrophoresis.
- A nylon filter or (nitrocellulose paper) is placed over it.
- Dry absorbent paper is used to draw the solution containing the fragments from the gel to the filter, leaving ‘blots’ on it.
- The alkaline solution also denatures the fragments so the strands separate and the base sequences are exposed.
Gene probes
- These are short DNA sequences that are complementary to specific sequences which are required.
- Hybridisation - large amounts of the probe are added to the filter and bind with the complementary DNA strands.
- Excess probes are washed away.
- X-ray pictures are taken of the filter or the filter is placed under UV light.
Polymerase chain reaction
The reactants:
- DNA sample
- DNA polymerase (enzyme)
- Primers
- The four nucleotide bases
- The reactants are placed in a vial in a PCR machine.
- Mixture is heated to 90-95°C – which causes the DNA strands to separate as the hydrogen bonds between them break down.
- Mixture is cooled to 50-60°C – the primers bind (anneal) to the single DNA strands.
- Mixture is heated to 75°C – the optimum temperature for the enzyme to build the complementary strands of DNA.
This is the end of the topic
Drafted by Eva (Biology)
Photo references:
- https://www.youtube.com/watch?v=YF0UaIB0QpE
- https://ib.bioninja.com.au/standard-level/topic-3-genetics/35-genetic-modification-and/gel-electrophoresis.html
- https://www.mun.ca/biology/scarr/Gr12-18.html
- https://zh.wikipedia.org/zh-hk/File:Polymerase_chain_reaction-en.svg