electrophoresis

Question 1

what is electrophoresis used for?

Answer 1

separating nucleotides

Question 2

step 1 (casting tray)

Answer 2

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

Question 3

step 2 (agarose gel)

Answer 3

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

Question 4

step 3 (wells)

Answer 4

• 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

Question 5

step 4 (load wells)

Answer 5

• 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

Question 6

step 5 (gel plate immersed in charged buffer solution)

Answer 6

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

Question 7

how do you complete a southern blot?