electrophoresis Flashcards

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1
Q

what is electrophoresis used for?

A

separating nucleotides

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2
Q

step 1 (casting tray)

A
  • glass plate has electrodes at each end (pos at one end, neg at other)
  • attached to electric supply
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3
Q

step 2 (agarose gel)

A
  • masking tape each side of tray
  • pour agarose onto glass plate
  • left to set/solidify
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4
Q

step 3 (wells)

A
  • use a comb to push down gel to form wells in the solidified agarose gel
  • at negative electrode end
  • add buffer to whole gel sheet to control pH which would affect hydrogen bonding
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5
Q

step 4 (load wells)

A
  • add a dye to the wells to allow DNA to become visible
  • DNA is broken into fragments using restriction endonucleases
  • add known DNA fragments to each well
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6
Q

step 5 (gel plate immersed in charged buffer solution)

A
  • connect to power supply
  • leave for 30 mins
  • smaller fragments move furthest and fastest
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7
Q

how do you complete a southern blot?

A
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