B. 1) Electrophoresis Flashcards
charged molecules moving through a matrix in an electric field will do what
move towards electrode with opposite charge
how is the rate of movement of molecules determined
by charged, size and shape. eg, movement of longer molecules slowed by friction
common use for electrophoresis
to determine the approx size of fragments of DNA and the size and subunit composition of proteins. the size can be determined by comparison of molecules of known molecular weight
DNA fragments, what ingredients are used
matrix used is made from agarose, a carbohydrate polymer isolated from algae. it is boiled and then cooled to form a gel. samples are added to wells formed at one end of gel
how will the DNA move
DNA -ve charge therefore movement through gel based on size
proteins, what ingredients are used
matrix made by the polymerization of polyacrylamide. a reducing agent like dithiothreitol (DTT) may be used to break disuphide bonds. and SDS-PAGE
what does SDS-PAGE stand for
sodium dodecyl sulfate polyacrylamide gel electrophoresis
what is SDS
-ively charged detergent. SDS denatures and coats proteins. combo of DDT and SDS denatures proteins, separates subunits and give them a uniform charge. movement of polypeptides through gel based on size