evidence for evolution Flashcards
genome
the complete set of genetic material in a cell; an organism’s complete set of DNA
DNA cloning
- researchers “clone” – make many copies of – a DNA fragment of interest, such as a gene
- involves inserting a target gene into a circular DNA molecule called a plasmid
PCR acronym
polymerase chain reaction
PCR function
small quantities of DNA to be replicated from a sample (amplified) → producing testable amounts to use in analysis techniques
what does development of PCR enable?
small quantities of DNA to be replicated from a sample → producing testable amounts to use in analysis techniques
- mimics the natural process of DNA replication that occurs prior to cell division
PCR steps
- denaturing
- annealing
- extension
how many times are the steps of PCR repeated?
sequence repeated 20-30 times in a process called thermocycling
thermocycling
process of repeated heating and cooling
how long does it take to produce a billion copies of DNA through PCR
2-3 hours
denaturing
- During natural DNA replication, the enzyme helicase separates the two strands of DNA (allows each strand to be copied)
- PCR uses heat to achieve same function
what temperature does denaturing occur at and why
- Temps of 94-96C used to break hydrogen bonds holding the two strands together
- Separates strands without disrupting each individual strand
annealing
- Allows short strands of DNA called primers to bind to the single DNA strands
- Primers not random sections of DNA → they’re complementary to either end of the section of DNA to be copied
what temperature does annealing occur at
Temperature is decreased to 50-60C
another name for extension
elongation
extension/elongation
- Mimics the process of DNA replication
- Enzyme DNA polymerase is used to join new, complementary nucleotides to the sections originating with the primers
- This extends the nucleotide chain → creates new strands of DNA
- Not the full length of original DNA – starts @ primer not @ end of DNA
- Eventually, majority of DNA strands are the length of DNA b/w location of the primers
- DNA polymerase attaches to double-stranded DNA
- Primers act as starting point → initiate DNA replication
what temperature does extension occur at?
temp increased to 72 degrees
all PCR applications now use heat stable DNA polymerase, what is this polymerase called?
Taq polymerase
where is taq polymerase taken from?
taken from a bacterial enzyme
why is taq polymerase more useful?
- Doesn’t denature when heated
- Allowed the procedure to be simplified and automated → sample can be alternately heated and cooled
how is PCR useful in the study of human evolution?
as it:
- provides testable amounts of DNA from very little
- eg. fossils – usually found in minute quantities
what other than the study of human evolution is PCR useful for?
- DNA profiling – comparing an individuals DNA to a library of DNA from known individuals
- early detection of infectious disease – foreign DNA from viruses/bacteria
- eg. COVID
- medical diagnostics
- forensic analysis
restriction enzymes
- Enzymes that cuts strands of DNA at specific sequence of nucleotides
what happens when restriction enzymes are added to DNA?
- When they are added to DNA → cuts strands into different lengths depending on base sequence of the specific DNA sample
- Length of pieces can be analysed and compared with other DNA samples
gel electrophoresis
- fragments of DNA are pulled through a gel matrix by an electric current by separating DNA fragments according to size
- pulls the negatively charged DNA through the gel from negative to positive electrode
- Technique that uses the banding patterns of DNA fragments as a means of identification; unique to every individual
banding pattern
individuals DNA profile or DNA fingerprint
process
- DNA fragments are placed in a gel
- Weak electric field is applied to the gel
- Electric field pulls the DNA fragments to one end of the gel/fragments move from the negative end (anode) to positive end (positive electrode)
- Fragments move @ diff. rates depending on size
which size fragments move the fastest in gel electrophoresis
Smallest fragments move faster
how are the fragments for different species separated?
in unique patterns
filling the wells
- the wells where the DNA is placed are simply depression in the gel
- DNA will rather move through the gel than diffuse through the solution
- DNA needs to be accurately placed in the wells using a micropipette (these reduce chances of cross contamination as they have disposable tips)
DNA ladder
- Often run at the same time as the samples
- Ladder contains segments of DNA with known lengths
- Results from unknown sample are compared to the ladder to determine the length of the DNA strands in the sample
what stains can be used for visualising DNA in gel electrophoresis
- ethidium bromide
- methylene blue
visualising DNA by using ethidium bromide
- As DNA moves through gel it picks up some of the chemical
- When run is completed, UV light is shone over gel and the DNA fluoresce
visualising DNA by using methylene blue
- Dye that binds to DNA
- When the gel is soaked in the dye, areas containing DNA stain a deeper blue → visible to naked eye
DNA sequencing purpose
Determining of the precise order of nucleotides in a sample of DNA
specific method used in DNA sequencing
Sanger sequencing
DNA (sanger) sequencing process
- The region of DNA to be sequenced is identified and cut (using bacterial enzymes)
- PCR amplifies the sample and creates dye labelled fragments.
- DNA sections are separated using gel electrophoresis (or other chromatography methods)
- Bands form which represent different sizes of DNA
what is used to sequence the DNA sample
computer software
uses for DNA sequencing (medicine)
- Used to identify mutations
- Compare DNA from different organisms
- Useful in identifying inherited disorders (sickle-cell anaemia, cystic fibrosis and cancer) and maternity/paternity tests
- Fertility
- Identify disease causing mutations based on family history
- Family planning
uses for DNA sequencing (science)
- Used by scientists to compare species in order to track evolutionary changes
uses for DNA sequencing (agriculture)
mapping and sequencing genomes of microorganisms making them useful for crops and food plants
uses for DNA sequencing (forensics)
to help identify individuals because each individual has a different genetic sequence
principles for ethical considerations with genetic information
- autonomy
- confidentiality
- equity
- privacy
autonomy
- Respect for the right to be self-determining; choose whether or not to be tested and if tested, to know and share the info
- Right of an individual to decide their own future, independent of genetic info
confidentiality
- Use of genetic info treated sensitively
- Accessed only by those who are authorised to access it
equity
The right to fair and equal treatment regardless of genetic info
privacy
- The right to be left alone
- Make decisions regarding genetic testing and the resulting information, independent of others
comparative studies
- genomics DNA
- genomics mtDNA
- proteins
- bioinformatics
- comparative genomics
- embryology
- homologous structures
- vestigial structures
genomics DNA
- DNA is universal; sequence of organic bases b/w species varies
- New genes gained by mutation
- Lost by natural selection or genetic drift
- Species that are more closely related → more sim.
- Non-coding sequences of bases in DNA (junk DNA) show more similarities with more closely related species → evolved from common ancestor
ERV acronym
endogenous retroviruses
what are endogenous retroviruses?
apparently non-functional DNA, a viral sequence that has become part of an organism’s genome
mitochondria
organelles in the cell where the aerobic phase of respiration occurs to release energy for use by the cell
what is mtDNA
Some DNA is located here (most in the nucleus) = mitochondrial DNA
In the form of small circular molecules (unlike strands of DNA in the nucleus)
- 5-10 molecules in each mitochondrion
how many genes in mitochrondrial DNA
37 genes = all essential for normal functioning
why is most cells containing large numbers of mitochondria an advantage?
- Lot easier to find and extract than DNA in the nucleus
- Smaller samples can be used