BIO505 Final Term Current Subjective

Principle of PCR and its reagents.

Principle of PCR

The principle of PCR (Polymerase Chain Reaction) involves the enzymatic amplification of a specific DNA sequence. It consists of repeated cycles of heating (denaturation), cooling (annealing), and DNA synthesis (extension). This process uses a DNA template, DNA primers, and a heat-stable DNA polymerase to exponentially produce copies of the target DNA region, facilitating various genetic analyses. Reagent use in PCR. Some of the key reagents used in a typical PCR reaction:

1. DNA Template: The starting DNA strand that contains the target sequence you want to amplify.
2. Primers: Short DNA sequences that define the start and end points of the target region to be amplified.
3. DNA Polymerase: The enzyme responsible for synthesizing a complementary DNA strand using the primers and nucleotides.
4. Nucleotides (dNTPs): The building blocks of DNA strands, containing adenine (A), thymine (T), cytosine (C), and guanine (G).
5. Buffer Solution: A solution that provides the optimal pH and salt concentration for the PCR reaction to occur.

PCR uses /Applications of PCR

PCR has applications in DNA amplification, Genetic testing, forensics, medical diagnostics, Environmental studies, Evolutionary studies, food safety, Archaeology, and anthropology

Which diseases are caused by x-linked disorder?

X-linked recessive diseases are genetic disorders caused by mutations in genes located on the X chromosome. Because females have two X chromosomes, they can be carriers of X-linked recessive mutations without showing symptoms. Males, who only have one X chromosome, are more likely to exhibit symptoms of Xlinked recessive diseases if they inherit a mutant allele. Examples of X-linked recessive diseases include hemophilia, Duchenne muscular dystrophy, Huntington’s Disease, and red-green color blindness.

Difference b/w genetic & environmental causes of variations

Variations in traits can be caused by both environmental and genetic factors. Environmental factors include things like diet, exposure to toxins, and physical activity, which can influence gene expression and affect traits. Genetic factors, on the other hand, involve variations in DNA sequences that are inherited from parents and can directly impact traits. It's important to note that in some cases variations are a result of a combination of both environmental and genetic influences.

Erythrocyte lysis In RNA/Erythrocyte extraction from RNA purification

Erythrocyte lysis is a process used to break open red blood cells (erythrocytes) in order to release their contents, including RNA. This process is commonly used in RNA extraction protocols to obtain RNA samples from whole blood, which contains a mixture of various cell types, including erythrocytes, leukocytes, and platelets. One commonly used method for erythrocyte lysis in RNA extraction is the use of a hypotonic buffer, such as ammonium chloride, which causes the erythrocytes to swell and burst, releasing their contents into the solution. The leukocytes and platelets are relatively resistant to lysis under these conditions, allowing for the isolation of RNA from these cell types. After erythrocyte lysis, the RNA can be extracted from the solution using various methods, such as column-based or bead-based purification. The extracted RNA can then be further purified and quantified for downstream applications, such as gene expression analysis or sequencing.

Procedure of western blotting

The Western blotting procedure involves several (6) steps. First, proteins are separated by size using gel electrophoresis. Then, the proteins are transferred onto a membrane. The membrane is blocked to prevent non-specific binding, and then incubated with a primary antibody that binds to the target protein. After washing, a