PCR
polymerase chain reaction
-create multiple copies of a specific section of DNA from a sample (amplification)
-useful when only small amounts of DNA available for analysis
Stages of PCR
Denaturing = heating DNA to separate the two strands
Hybridisation/Annealing = primer added to DNA & bind to complementary base sequences on separated strands (starting point for replication)
synthesis/Elongation = DNA polymerase synthesis DNA code and build complementary strand of DNA (34 min)
Thermocycling
process of repeated heating and cooling
Gel electrophoresis
technique - separate DNA strands based on their lengths
Process of gel electrophoresis
-DNA pieces placed in wells in semi-solid gel - immersed in solution of an electrolyte
-electrodes at either end of gel (negative electrode close to DNA & positive opposite side)
-electric current passed through gel, DNA (- charge) move towards + electrode
-small DNA pieces move faster = located further away from negative electrode when current stopped
DNA profile
banding pattern on gel electrophoresis
-used to establish individual’s identity/parentage
Filling the wells
-wells = depressions in gel
-DNA accurately placed in wells using micropipette (disposable tips can be put on n off without contact = ↓ chance of cross-contamination)
DNA ladders
-run at the same time as samples
-contain segments of DNA with known lengths = compare unknown sample to determine length of DNA strands in sample
Visualising DNA
-ethidium bromide added to agar prior to get being set = DNA picks up chemical
-DNA fluoresce under special uv light
-methylene blue = dye binds to DNA
-areas containing DNA stain = a deeper blue = visible to naked eye
DNA probes = short sections of a single strand of DNA with. radioactive or fluorescent molecule - binds to DNA tested
DNA sequencing
determination of precise order of nucleotides in a sample of DNA
-identify mutations/ compare DNA from different organisms
-useful in identifying inherited disorders = sickle cell anaemia, cystic fibrosis + some cancer
-maternity and paternity tests
-compare species - track evolutionary changes
When DNA forms
each nucleotide loses two phosphate groups
sugar molecule loses a Hydrogen atom from OH group (hydroxy) when bond to phosphate group of an adjacent nucleotide
Sanger method
-nucleotides that lack OH group added to growing strand (ddNTPS = dideoxyribonucleotides/ dideoxyribonucleotide triphosphate)
-stop elongation of sequence because no OH group for next nucleotide to attach to = create different lengths of DNA
-separated (gel electrophoresis) - know which base was added - create each length length = determine order of nucleotides
Autonomy
-respect for right to be self-determining & choose whether or not to be tested
-right of individual to decide their own future, independent of genetic information
Confidentiality
use of genetic information is treated sensitively, is accessed only by those who are authorised to access it
Equity
right to fair and equal treatment regardless of genetic information
Privacy
right to be ‘left alone’ and to make decisions regarding genetic testing & resulting information, independent of others
evolution
gradual process of change in the inherited traits of a population of organisms from one gen to the next
-permanent change in a population’s gene pool from one gen to the next
Fossil record
organisms in the past not same as organism alive today (phylogeny of the horse)
Amino acid sequencing
-proteins = long sequences of amino acids
-sequences very similar = closely related species
-sequence less similar = distantly related species
-indicate evolution from common ancestor
Ubiquitous proteins
-found n all living organisms
e.g. Cytochrome C = protein used to produce energy
-contain 104 amino acids = strong evolution for ancestral evolution of all species
Comparative studies of DNA
-sequences of bases (AT,G,C) similar in closely related species
-sequence less similar = more distantly related species
-similarity of DNA indicates evolution from a common ancestor
Mitochondrial DNA (mtDNA)
-small amt of DNA present in mitochondria )small & circular with 37 genes needed of mitochondrial function)
-mtDNA extraction easier than nucleic DNA
-only inherited from mother’s egg
-mutation rate of mtDNA high,
Bioinformatics
involves use of computers (software, statistics, mathematics & engineering) = describe molecular components of living things
-trace evolution of organisms by looking at changes in DNA
Annotation
whole genomes have been compared in a process
Mineral replacement
-varying degrees of mineral replacement (0-100%)
Stage 1 - minerals from groundwater fill pores in the bone or shell (permineralisation)
Stage 2 -minerals forming the bone matrix/shell - dissolved away and replaced by minerals in groundwater
carbonisation
occurs in very fine-grained rocks
mummification
occurs when soft and/or hard tissues exposed to chemicals, extreme cold, very low humidity, or lack of air (bodies sealed in fine sediment, resin or tar)
Relative dating
-compare age of one thing with that of something else
Superposition
lower beds in a sedimentary sequence are older than rocks above
Youngest
↑
|
↓
Oldest
Comparative stratigraphy
study of sedimentary rocks
-sequence of sedimentary rocks in different areas is similar = likely are of same age
index fossil
fossil of a species that can be used for relative dating
-distinctive appearance, short time span + have broad geographical distribution
Radiometric dating
absolute dating method for determining chronological age of a rock or mineral by measuring proportions of original radioactive material and its decay product
Half life
unique rate of decay - time taken for half of any given amount of isotope to decay
Common radioisotopes commonly used for radiometric dating
Carbon-14 → Nitrogen-14
Potassium-40 → Argon-40
Limitations of radiometric dating methods
C14 dating - must be organic (has carbon), maximum is 50000 - 60,000 years
Potassium-argon dating must be igneous rock, minimum age 200,000 years