gel electrophoresis Flashcards
why do people do it
to sort and measure DNA strands according to lengths
why is DNA negatively charged
because of the phosphate groups in the backbone of the molecule
how does dna move through gel
electric current is passed through gel, since DNA is negatively charged, the fragments are repelled by a negative charge attracted by a positive charge
procedure
1
RE cut DNA into fragments
procedure
2
DNA sample is placed in a well at the top of a gel.
gel is porous- it has tiny holes so that dna fragments can squeeze through
procedure 3
electric current is passed through.
smaller fragments can squeeze through more easily, so they move the fastest and furthest along the gel (larger fragments are opposite).
procedure 4
DNA ends up forming bands in the gel at different locations, depending on the different locations, depending on the size and number of fragments
how to know fragment size
is determined by “running” a known DNA sample beside well so the band locations can be compared.
results of the know sample creates a “ladder” with bands at known lengths measured in bp
how to make it visible
ethidium bromide under UV light
- simplest method is to dye the DNA with a molecule that will attach to the DNA and can be seen.