Manipulating genomes Flashcards
1
Q
Genome
A
All the genetic material contained by an organism
2
Q
DNA profiling
A
- Extract the DNA from tissue sample produced by PCR
- Digesting the sample using restriction endonuclease
- Separating the DNA by electrophoresis
- Hybridisation (radioactive or fluorescent probes added to excess DNA in order to label the fragments)
- Development (membrane with radioactively labelled fragments is placed onto X-ray film) - this reveals dark bands where the radioactive fluorescent DNA probes are attached.
3
Q
PCR
A
- Strands are separated at 90-95 degrees for 30 seconds, which denatures the DNA by breaking the H bonds.
- Annealing the strands by adding primers to the ends of the DNA strands at 55 degrees. Needed for replication of DNA to occur.
- Synthesis of DNA as temp is increased to 72-75 degrees for 1 min, so DNA polymerase can work best and add bases to the strands. Two new DNA fragments are produced.
4
Q
Uses of DNA profiling
A
- criminal investigations
- forensic science
- proof of paternity
- checking if individuals are prone to certain diseases
5
Q
Sanger sequencing/DNA sequencing
A
- DNA is placed into 4 tubes and primers, DNA polymerase, and activated free nucleotides are added.
- In each tube, chain-terminating nucleotides for one specific base are added.
- Tubes are placed in a THERMOCYCLER and PCR begins.
- PCR occurs until the DNA polymerase reached the CTN, where PCR stops.
- The terminator base attaches at different lengths down the sequence, therefore, DNA strands with different lengths are produced.
- Capillary sequencing (like gel electrophoresis) is used to separate them according to their length.
- The end of each DNA strand can be identified by the fluorescent markers.
- Order of bases in the capillary tube determines the sequence of the new DNA strand
6
Q
Bioinformatics
A
Development of the software and computing tools needed to organise and analyse raw biological data.
7
Q
Computational biology
A
Uses the data provided by bioinformatics to build theoretical models of biological systems, used to make predictions in certain circumstances.
8
Q
Spliceosomes
A
- mRNA includes exons and introns, and introns have to be removed.
- exons to be translated are joint together by spliceosomes
