Dna Electrophoresis Lab
DNA Extraction from Wheat Germ and Making an Agarose Gel
AIM:
To be able to make and agarose gel and perform gel electrophoresis in six different dyes. Also, to extract DNA from wheat germ.
INTRODUCTION:
Agarose gel is a substance that is used in science for gel electrophoresis and size exclusion chromatography. These processes use agarose gel to separate and analyze proteins and DNA. The medium is composed of a purified agarose powder that has been boiled in a buffer solution and then cooled into a gel. Agarose gel is most commonly associated with gel electrophoresis. In this procedure, scientists use an electrical charge to move deoxyribonucleic acid, more commonly known as DNA, or ribonucleic acid (RNA) through a gel matrix toward a positive pole. Because the molecules have to move through small holes in the lattice bonds in the agarose gel, smaller molecules move much faster than larger molecules. Using ultraviolet imaging of the molecules' movement and a formula that relates molecular weight to the speed of travel through the gel, scientists can determine the size of the molecules. Electrophoresis is essentially a sieving process. The larger the fragment of DNA, the more easily will it become entangled in the matrix and, therefore, the more slowly will it migrate. Small fragments, therefore, run more quickly than large fragments at a rate proportional to their size. The relationship of size to migration rate is linear throughout most of the gel, except for the very largest fragments. Large fragments have a more difficult time penetrating the gel and their migration, therefore, does not have a linear relationship to size. The matrix can be adjusted, though, by increasing the concentration of agarose (tighter matrix) or by decreasing it (looser matrix). A standard 1% agarose gel can resolve DNA from 0.2 - 30 kb in length. Agarose is extracted in the form of agar from several species of red marine algae, or seaweed, found in California and
AIM:
To be able to make and agarose gel and perform gel electrophoresis in six different dyes. Also, to extract DNA from wheat germ.
INTRODUCTION:
Agarose gel is a substance that is used in science for gel electrophoresis and size exclusion chromatography. These processes use agarose gel to separate and analyze proteins and DNA. The medium is composed of a purified agarose powder that has been boiled in a buffer solution and then cooled into a gel. Agarose gel is most commonly associated with gel electrophoresis. In this procedure, scientists use an electrical charge to move deoxyribonucleic acid, more commonly known as DNA, or ribonucleic acid (RNA) through a gel matrix toward a positive pole. Because the molecules have to move through small holes in the lattice bonds in the agarose gel, smaller molecules move much faster than larger molecules. Using ultraviolet imaging of the molecules' movement and a formula that relates molecular weight to the speed of travel through the gel, scientists can determine the size of the molecules. Electrophoresis is essentially a sieving process. The larger the fragment of DNA, the more easily will it become entangled in the matrix and, therefore, the more slowly will it migrate. Small fragments, therefore, run more quickly than large fragments at a rate proportional to their size. The relationship of size to migration rate is linear throughout most of the gel, except for the very largest fragments. Large fragments have a more difficult time penetrating the gel and their migration, therefore, does not have a linear relationship to size. The matrix can be adjusted, though, by increasing the concentration of agarose (tighter matrix) or by decreasing it (looser matrix). A standard 1% agarose gel can resolve DNA from 0.2 - 30 kb in length. Agarose is extracted in the form of agar from several species of red marine algae, or seaweed, found in California and