Electrophoretic Techniques

Question 1

advantages of EP?

Answer 1

high res, sensitive, multiplicity, simple, cheap

Question 2

types of EP systems

Answer 2

vertical (PAAG, rods, slabs)

horizontal (agarose, NAs)

Question 3

role of gel matrix

Answer 3

inhibits convection and diffusion, neutral, provides additional separation

Question 4

agarose gel structure

Answer 4

repeating unit agarobiose, galactose, anhydrogalactose

H bonding

Question 5

what is resolution

Answer 5

separation relative to band width

Question 6

what effects resolution?

Answer 6

sample vol and conc, time, gel density, mode

Question 7

why are proteins fixed in methanol before EP

Answer 7

to denature and ppt proteins to prevent diffusion

Question 8

what do horizontal streaks in 2D PAGE mean

Answer 8

ionic impurities or insuf IEF

Question 9

what do vertical streaks in 2D PAGE mean

Answer 9

insuf equil or not enough SDS

Question 10

western blot technique

Answer 10

protein blot, selective staining, mAB

Question 11

DNA blot

Answer 11

southern blot - cleave DNA with RE, hybridization probe

Question 12

RNA blot

Answer 12

northern, hybridization probe