Molecular Techniques
Molecular techniques are methods employed in molecular biology, genetics, biochemistry, and biophysics to manipulate and analyze nucleic acids (deoxyribonucleic acid (DNA) and ribonucleic acid (RNA)), protein, and lipids. Techniques in molecular biology are employed to investigate the molecular basis for biological activity. These techniques are used to analyze cellular properties, structures, and chemical reactions, with a focus on how certain molecules regulate cellular reactions and growth.
DNA Fingerprinting and Gel Electrophoresis
The genetic makeup of living organisms is shown by a technique known as DNA fingerprinting. The difference is the satellite region of DNA is shown by this process. Alex Jeffreys has invented the process of DNA fingerprinting in 1985. Any biological samples such as blood, hair, saliva, semen can be used for DNA fingerprinting. DNA fingerprinting is also known as DNA profiling or molecular fingerprinting.
Molecular Markers
A known DNA sequence or gene sequence is present on a chromosome, and it is associated with a specific trait or character. It is mainly used as a genetic marker of the molecular marker. The first genetic map was done in a fruit fly, using genes as the first marker. In two categories, molecular markers are classified, classical marker and a DNA marker. A molecular marker is also known as a genetic marker.
DNA Sequencing
The most important feature of DNA (deoxyribonucleic acid) molecules are nucleotide sequences and the identification of genes and their activities. This the reason why scientists have been working to determine the sequences of pieces of DNA covered under the genomic field. The primary objective of the Human Genome Project was to determine the nucleotide sequence of the entire human nuclear genome. DNA sequencing selectively eliminates the introns leading to only exome sequencing that allows proteins coding.
Procedure
- 1 to 2% of agarose gel was prepared in the running buffer. Mixture was swirled and mixed. The agarose and buffer mixture were boiled and mixed well until the agarose was completely dissolved.
- The agarose was cooled on the benchtop or by incubation in a 65 °C water bath.
- The gel tray was placed into the casting apparatus. The appropriate comb was placed to create the wells. The molten agarose was poured into the gel mold.
- The agarose was allowed to set at room temperature. The comb was removed and the gel was placed in the gel box. The running buffer was added until it covered the surface of the gel. A loading dye was loaded and the serum protein samples to be separated carefully.
- The lid was replaced to the gel box. Ensure the leads were plugged into the correct slots in the power supply. The power was turned on and started the electrophoresis running until the dye was migrated to an appropriate distance.
- After electrophoresis, the gel was stained with Coomassie brilliant blue and detain with detaining solution.
Question : Label albumin, alpha 1, alpha 2, beta and gamma on the result part
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