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DNA extraction (source of DNA (Plants (three types of DNA (genomic DNA,…
DNA extraction
source of DNA
Plants
three types of DNA
genomic DNA
mitochondria DNA
chloroplast DNA
example : fern, oil palm
cell support : cell wall
Animals
two types of DNA
genomic DNA
mitochondria DNA
cell support : cell membrane
example: ants, sheep
Prokaryotes
two types of DNA
genomic DNA
plasmid DNA
Cell support : peptidoglycan
example: bacteria
3 major step in bacterial
chloroform extraction
mixing is achieved through inversion of microcentifuge tube several times.
additional of equal volume of chloroform to cell lysate
apply centrifuge to separate the phases
only retrieve the aquoes phase for the next stage of work
lysis of bacterial cell
vortex
will produce uniform consistency throughout the solutions
vortex machine
to mix fluid through very fast spins
centrifuge
will produce layers or sedimentation of pellets
centrifuge machine
to separate substances according to their densities by application of centrifugal force
DNA precipitation
mix the components through inversion of microcentifuge several time
facilitated better DNA precepitation by placing the tube on ice at least 30'
additional of 2 volumes of ice-cold absolute ethanol to the clear cell lysate
DNA will precipitate into ethanol as cloudy materials.
Resuspension of pellet DNA
place the tube on ice ~15 minutes to faciliated
store the DNA in ~20 minutes
add sterile distilled water or TE buffer to pellet DNA