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.
Question. What is the major difference between the conventional PCR and real time PCR (qPCR) in terms of function?
Answer : Major differences between the conventional PCR and real time PCR -
Cnventional PCR : Conventional PCR is not quantitative , but rather qualitative. In conventional PCR , the amplified DNA product, or amplication, is detected in an end - point analysis. Conventional PCR is not ideal for rapid diagnostic use in most clinical laboratories. It has been used to detect , monitor and identify fungi from an entire set of environmental samples and is the core of molecular fungal diagnostics. The conventional PCR is based on the three simple steps required for any DNA synthesis reaction : 1. denaturation of the template into single strands; 2. annealing of primers to each original strand for new strand synthesis; and 3. extention of the new DNA strands for the primers.
Real time PCR : also called quantitative PCR (qPCR), and it is an automated technique that allow for detection and quantification of the target material. RT-PCR is a method frequently used to screen large cell populations , and provide rapid genotyping by HRM. IN real time PCR , the accumulation of amplification product is measured as the reaction progresses , in real time, with product quantitification after each cycle.
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