electrophoresis Flashcards
product of PCR
amplicons
the movement of molecules by (DNA or RNA) by (applying specific voltage) an electric current
electrophoresis
electrophoresis detect and quantitiate —
DNA or nucleic acid
electrophoresis can occur in —
air or solution or in a matrix to limit migration and contain the migrating material
electrophoresis is routinely applied to the —
analysis of nucleic acids/proteins molecules
mostly negative charged
Each phosphate group on a nucleic acid polymer is ionized making the molecule positively charged.
true or false
FALSE
negatively charged
Under an electric current, DNA and RNA will migrate toward the —
positive pole (anode)
The nucleic acid will move toward the positive pole (anode) because nucleic acids are negatively charged.
In a matrix of — migration under the pull of the current is impeded, depending on the size of the molecules and the spaces in the gel matrix
agarose or polyacrylamide
It is affected by the size and the charge of the particle.
origin or the point of application of electrophoresis
DNA and RNA becomes negatively charged
gel system provide resistance to the movement of molecules under the force of an electric current preventing — and reduce — so that the separated molecules from a defined group, or “band”
prevent diffusion and reduce convection currents
serve as a support medium for analysis of the separated components.
gel
agarose gel
size of the DNA –> concentration
high concentration: —
low concentration: —
high: impede migration
low: weak gel
true or false
small pieces of DNA (50-500 base pairs) are resolved on lower agarose gel, for example, 0.5% - 1%
FALSE.
resolved on more concentrated agarose gels
larger fragments of DNA (2,000 to 50,000) are best resolved in —
lower agarose concentrations
(0.5% - 1%)
The gel strength of any concentration of agarose will also
decrease over time and with exposure to chaotropic agents
such as urea.
drawback of agarose gel
high concentration will impede migration
low concentration produce a weak gel that is easily broken
gel suitable for very large pieces (50,000 to 250,000 + bp) of DNA
pulses of current applied to the gel in alternating
dimensions enhance migration.
pulse-field gel electrophoresis
pulse-field gel electrophoresis is used for —
bacterial typing for epidemiological purposes
this works by alternating the positive and negative
electrodes during electrophoresis. in this type of separation, the DNA goes periodically forward and backward
field-inversion gel electrophoresis (FIGE)
used for applications that require the resolution of chromosome-sized fragments of DNA, such as in bacterial typing for epidemiological purposes.
Alternating-field electrophoresis
this gel require a catalysts (APS + TEMED Light activation and in higher resolution capability for smaller fragments
polyacrylamide gels
