POLYACRYLAMIDE GEL

ELECTROPHORESIS

PAGE
POLYACRYLAMIDE
GEL
ELECTROPHORESIS(OR
PAGE)
DESCRIBES A TECHNIQUE TO SEPARATE
BIOLOGICALMACROMOLECULES,
USUALLYPROTEINSORNUCLEIC ACIDS,
ACCORDING
TO
THEIRELECTROPHORETIC
MOBILITY
USINGPOLYACRYLAMIDEGEL.
ELECTROPHORETIC MOBILITY IS A
FUNCTION
OF
THE
LENGTH,
CONFORMATION AND CHARGE OF THE
MOLECULE.

PAGE FOR DNA

DNAGELELECTROPHORESISISUSUALLY
PERFORMED
FOR
ANALYTICAL
PURPOSES,
OFTEN
AFTER
AMPLIFICATION OF DNA VIAPCR, BUT
MAY BE USED AS A PREPARATIVE
TECHNIQUE PRIOR TO FURTHER
CHARACTERIZATION.
FORNUCLEICACIDS,THEDIRECTIONOF
MIGRATION,
FROM
NEGATIVE TO
POSITIVE ELECTRODES, IS DUE TO THE
NATURALLY OCCURRING
NEGATIVE
CHARGE CARRIED BY THEIRSUGARPHOSPHATEBACKBONE.

POLYACRYLAMIDE GEL
POLYACRYLAMIDEGELIS
MADEBYCHEMICALCROSSLINKINGOFACRYLAMIDE
ANDCROSSLINKER(USUALLY
BIS-ACRYLAMIDE),
PRODUCINGAMOLECULAR
SIEVE.
ASOURCEOFFREE
RADICALSANDASTABILIZER,
SUCHASAMMONIUM
PERSULFATE ANDTEMEDAR
EADDEDTOINITIATE
POLYMERIZATION.

PROPERTIES OF POLYACRYLAMIDE
HIGH RESOLUTION FOR SMALL DNA
FRAGMENTS
WHILEAGAROSEISUSEDFORDNA
FRAGMENTSOF50-20000BPINSIZE,
POLYACRYLAMIDEGELSHAVEHIGH
RESOLVINGPOWERFORSMALL5-500BP
FRAGMENTSOFDNAANDAREALSO
GOODFORANALYZINGSINGLESTRANDEDDNA.
POLYACRYLAMIDEGELSCANSEPARATE
DNATHATDIFFERSBY0.2%INLENGTH,
WELLBEYONDTHERESOLVING
CAPABILITIESOFAGAROSE(2%
DIFFERENCEINDNALENGTH).

PROPERTIES OF POLYACRYLAMIDE
UNIFORM PORE SIZES
UNLIKEAGAROSEGEL,THEPORESIZE
OFPAGELSAREUNIFORMANDSMALL
ANDCANBEMADESMALLERBY
ADDITIONALCROSS-LINKING.
CONTROLOFTHEPORESIZEOFA
POLYACRYLAMIDEGELISACCOMPLISHED
BYCHANGINGTHET(TOTAL%W/V
ACRYLAMIDEANDCROSSLINKERINGEL)
ANDC(%W/VCROSSLINKERINGEL)
VALUES.THE MORE CROSSLINKS THERE
ARE, THE SMALLER THE PORE SIZE.

PROPERTIES OF POLYACRYLAMIDE
LONGER SETTING TIME, TOXIC, BUT MORE
CONSISTENT AND STRONGER
AGAROSEGELSETSPHYSICALLYQUICKLY
WHILEPAGELSETSCHEMICALLYBY
CROSSLINKINGWHICHHAPPENSSLOWER.
LOW%AGAROSEGELSAREWEAKWHILE
HIGH%AGAROSEGELSAREBRITTLE.
POLYACRYLAMIDEAREVERYSTABLEAND
CANBECASTASTHINGELSTHATPROMOTE
BETTERSEPARATION.
ACRYLAMIDEISAPOTENTNEUROTOXIN.
DISPOSABLEGLOVESMUSTBEUSEDWHEN
HANDLING.