Nucleic Acid Lab Report

Nucleic acids

Nucleic acid were discovered by Friedrich Miescher in 1869. They are generated within laboratory using enzymes and by solid-phase chemical synthesis. It is highly complex, long-chain polynucleotide mega biomolecules .DNA and RNA are the two nucleic acids. DNA play role as the genetic material while RNA is non-genetic. both RNA and DNA are made from monomers known as Nucleotides .each nucleotides has 3 components a 5-carbon sugar, a phosphate group and nitrogen base. If the sugar is ribose then it is RNA. While nucleosides is the substructure consisting of a nucleobase plus sugar

1-Lyse-Using lysis procedure the cells of a sample are broken down.

2-Bind –Along with ethanol or isopropanol a buffer solution is added to sample forming binding solution.the column is put in centrifuge while binding solution transferred to a spin column.inside the spin column centrifuge forses the binding solution through a silica gel membrane.the nucleic acid bind to the membrane as solution passes through when the pH and salt concentration of binding solution are optimal.

3-Wash-Wash buffer is added to column by removing flow-through.the column is put in centrifuge again,forcing the buffer through membrane,leaving only nucleic acid bound to the silica gel by washing.

4-Elute-BY removing the wash buffer elute is added to column is put in centrifuge again, forced the buffer through membrane.from the membrane elution buffer allows to removes nucleic acid and collected from bottom of the column.

BOOM METHOD-

1- It is solid phase extraction method for isolating nucleic acid from biological

► 2.5-5% is messenger RNA

RNA analysis-

Electrophoresis-

1-Nucleic acids are are negatively charged,which makes them migrate to the positive pole in the electric field.

2-The separation process depends on the voltage used,the properties of the gels,as well as the charge and shape of the molecule.

3-Agarose gel electrophoresis and polyacrylamide gel electrophoresis used for high resolution.

Agarose gel electrophoresis-

1 - it is used to separate DNA fragments that vary in size .Agarose is a polysaccharides obtsained from marine red algae .it is added to an electrophoresis buffer and then dissolved by heating it.

2 - The presence of many hydroxyl ngroup enable hydrogen brides to form,which lends firmness to the large pored gel matrix.

3 - The agarose gel is kepy in horizontal position and completely covered by the buffer which prevent it from drying out.

4- The speed at which DNA fragments migrate through agarose gels in the electric field depends on the size of the DNA fragments.

5- The migration speed of linear double-stranded DNA molecules is inversely proportional to the logarithm of their size