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Biotechnology techniques - Assignment Example

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Short Answers (Approximately 3 pages) 1. Why do you first denature a plasmid DNA template or a PCR product being used for sequencing? (10 points)  The plasmid DNA is denatured to convert the double stranded DNA molecule into single stranded DNA. This single stranded DNA molecule acts as a template for the sequencing reaction…
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Is this statement TRUE or FALSE, explain? (5 points) Electroporation is used to introduce DNA in prokaryotic and eukaryotic cells. This is the general method of introducing the foreign DNA into the desired cells. This method uses the electrical pulses to create a pore in the cell membrane and through these pores the foreign DNA molecule is inserted. 3. What are the 3 main sources of Stem Cells? (10 points) The 3 main sources of stem cells included Bone marrow, Bloodstream or peripheral blood and finally umbilical cord blood from the newborns.

Bone marrow particularly those in the pelvis’ bones have rice supply. Blood normally has few stem cells, and when hormone like substances called growth factors are given to individuals, stems cells grow faster and move from the marrow to the blood. Blood left in the placenta and the umbilical cord of the newborns can be retrieved, store and utilized for future.  4. What is meant by humanization of mouse antibodies and what is the need for "humanization"? (10 points) While using therapeutic antibodies or murine antibodies isolated from mice in humans, immunogencity problem arose due to the differing protein sequence.

To alleviate this issue, specific regions within a mouse antibody called complementarity determining regions (CDR) are identified and transferred into human variable regions and this process is called humanization. When this is done, it will confer binding and so the resultant humanized antibodies can function without major immunogencity problem.  5. Which technology cannot be used to determine the function of a gene? Explain. (5 points) Stem cell technology cannot be used to study the gene function because, it is a method to produce the entire cell and study the organs and it is not the direct method of analysis of gene function.   6. cDNA microarray technology specifically evaluates the gene expression levels of ONLY microRNA in cells and tissues.

Is this statement TRUE or FALSE? Explain? (5 points) No cDNA microarray technology is used for the analysis of all the genes and its function. cDNA technology is used to analyse the functions of known and unknown gene.   7. Which of the following gene expression strategies increase the number of different proteins available to the cell without increasing the total number of genes encoded by the genome?(5 points) Translation is used to increase the number of different proteins in the cell without increasing the total number of genes in the genome.

By varying the mRNA translation, the protein synthesis is modified.   8. In the sequence given below, how many EcoRI recognition sites are present? Please circle them. (10 points)      5’-ATGGAATTCCCAATTGAATTCCGGCTGAATTCAAA-3’     3’-TACCTTAAGGGTTAACTTAAGGCCGACTTAAGTTT-5’ There are three EcoRI recognition sites in the given sequences. They are  5’-ATGGAATTCCCAATTGAATTCCGGCTGAATTCAAA-3’     3’-TACCTTAAGGGTTAACTTAAGGCCGACTTAAGTTT-5’ 9. What is meant by the statement, "Complementation screening of genomic eukaryotic clones will not work in E.

coli because of absence of splicing machinery?" (10 points) E.coli is a prokaryote. The bacterial systems cannot remove the introns from the mRNA sequences. Complementation screening is the technique used to identify the mutations in the gene of interest. In this method, the selection is based on the capacity of the gene to get back the phenotype. The splicing mechanism is not present in the

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