How many chemical base pairs are in the human genome?
The human genome is composed of 3 billion base pairs located in the 23 chromosomes and is found within the cellular nucleus. These nucleic bases include Adenine, thymine, cytosine, and guanine in a DNA molecule and for RNA the thymine is replaced with Uracil.
What is the purpose of PCR?
Polymerase chain reaction is a molecular technique always used to make billions of copies of a given segment of DNA. It is mostly used to amplify desired DNA segments, which can then be used for other purposes such as cloning, diagnosis, sequencing among others.
What is the function of the primer
Artificially made pieces of DNA that are single-stranded contains 20 to 30 nucleotides and must be present for DNA polymerase to synthesize a new strand. Primers often anneal to the two separated DNA strands and form the basis of amplification as they determine the nucleotide sequence to be added onto the new strand.
What are nucleotides?
Nucleotides are organic molecules that act as the building block for nucleic acid. DNA molecule is made up of many nucleotides joined together with a bond. Nucleotides consist of a sugar molecule, one phosphate group/molecule and one of the four bases (ACTG).
What is the last step of the experiment?
DNA Extension: It involves the synthesis of new DNA strands. In this step, the temperature within the thermocycler is slightly increased to 72 degrees Celsius, Taq polymerase enzyme then adds dNTPs, deoxyribonucleotide triphosphate onto the new strand from 5' to 3' resulting in the formation of a new daughter strand (Ahrberg et al., 2016).
What is the DNA thermal cycler?
DNA thermal cycler is a laboratory apparatus/instrument used to amplify sections of DNA molecule during the PCR process. It functions by changing internal temperature to allow for different steps of PCR to set in, thereby allowing amplification of the required DNA segments.
What happens at 203F?
At 203F, the double-stranded DNA molecule is denatured. It is characterized by separation of the Helical strands of DNA into two single strands thereby allowing primers to anneal to the separated strands and latter amplification occurs.
What happens at 122F?
At 122F, the forward and reverse primers anneal to the two denatured strands of DNA molecule. The annealing occurs at specific sequences along the two strands. The sequences on which the primers anneal often have complementary bases to those of the annealing primers.
What happens at 72C?
It is the optimal operating temperature at which Taq polymerase enzyme functions best. At this temperature, Taq polymerase attaches onto the primers and adds DNA nucleotides onto the new strand thus extends/ synthesizes a daughter strand.
In cycle 2, why is the temperature raised?
At the second cycle, the temperature of the thermocycler is often raised to initiate the breakdown of the double-stranded DNA molecule into two single strands. This, therefore, allows primers to anneal onto the two strands and Taq polymerase to initiate the replication process.
What are the desired products of cycle 3?
At cycle three, Target sequence defined by the primers begins to accumulate.
After 30 cycles, what is your product?
Many copies approximately a billion copies of the sequence in question are produced from a single beginning molecule. Therefore, this indicates a successful replication of the desired DNA sequence/segement.
You are a researcher interested in studying genes that have an increased level of activity in skin cancer cells, what color spots should you choose?
To detect an increased level of activity of a gene in skin cancer cell, Red acts as the best color spot to be selected.
What is the job of gene4263?
Gene 4263 is responsible for Production of a protein whose function is to turn down expression of several other genes. This often acts as gene regulators.
What color are the spots regulated by gene4263?
Green is often the color regulated by the gene 4263.
What color is represented by 6219?
The color Yellow often represents the gene 6219.
What does RNA expression usually indicate?
RNA expression often indicates genes, which are actively expressed in a given cell/organ.
Why is gene6219 the color it is?
Gene 6219 is often yellow in color because it contains genes hybridized to both green and red DNA.
Why can't 6219 be detected in the microarray analysis?
The microarray cannot detect gene 6219, because the genes are combined with both the green and red DNA.
List 3 things microarray analysis cannot do?
Microarray analysis is mostly used to detect gene expression. However, the technique cannot identify splice variance, it cannot analyze the DNA molecule, which is not hybridized, and it cannot analyze sequences that are not predefined.
Ahrberg, C. D., Manz, A., & Chung, B. G. (2016). Polymerase chain reaction in microfluidic devices. Lab on a Chip, 16(20), 3866-3884.
Nguyen, H. T., Dupont, L. N., Jean, A. M., Gehin, T., Chevolot, Y., Laurenceau, E., & Gijs, M. A. M. (2018). Microfluidic extraction and microarray detection of biomarkers from cancer tissue slides. Journal of Micromechanics and Microengineering, 28(3), 034004.
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