strong light (e.g. blue light) and emits fluorescent signal which is proportional to the concentration of DNA in the gel (Thermofisher Scientific‚ 2011). In this experiment‚ SYBRSafe is used for the detection of mutant OFP gene and the corresponding Plasmid DNA‚ pRSET vector after restriction digestion by KpnI and HindIII. Signal strength of SYBRSafe can be one of the parameters to measure the efficiency of restriction digestion. Q2. What is the purpose of the addition of ampicillin to the LB-agar plates
Premium DNA
insert a gene into a plasmid. Someone gives you a preparation of genomic DNA that has been cut with restriction enzyme X. The gene you wish to insert has sites on both ends for cutting by restriction enzyme Y. You have a plasmid with a single site for Y‚ but not for X. Your strategy should be to A) insert the fragments cut with X directly into the plasmid without cutting the plasmid. B) cut the plasmid with restriction enzyme X and insert the fragments cut with Y into the plasmid. C) cut the DNA
Premium DNA
differ mainly in their plasmids. Zakharyan R.A et al. first reported the presence of plasmids in B. thuringiensis and suggested involvement of the plasmids in endospore/crystal formation. They also described the presence of large plasmid in the Cry+ variant of B. thuringiensis Upon sporulation‚ B. thuringiensis forms crystals of proteinaceous insecticidal δ-endotoxins (Cry toxins) which are encoded by cry genes‚. It was determined that the "cry" genes are harbored in the plasmids in most strains of
Premium Bacillus thuringiensis Insect Bacteria
to control DNA through the use of plasmid vectors became an attraction for newer technologies‚ which allowed specific changes in discrete‚ manageable segments of the genome with relatively little effort. Site-directed mutagenesis method was first benefited from recombinant DNA technology in 1970s‚ when isolated genes were exposed to conditions such as chemical agents or nucleotide analogs to localize their mutagenic effects. During this time‚ the use of plasmid vectors for DNA replication greatly
Premium DNA
Harrison E‚ Koufopanou V‚ Burt A‚ MacLean RC. 2012. The cost of copy number in a selfish genetic element: the 2-μm plasmid of Saccharomyces cerevisiae. J Evol Biol 25(11): 2348-56. What are the actors (e.g.‚ what parts of the organism are in conflict) and what are they in conflict over? The 2 μm plasmid of Saccharomyces yeast is in conflict with the cell host‚ this plasmid cost the host through using the cells’ resources ; meaning a burden on the host to synthesize more proteins as well as
Premium Gene DNA Molecular biology
* State the biotechnology is the industrial use of living organisms to produce food‚ drugs of other products * Explain why microorganisms are used in biotechnological processes * Describe the standard growth curve in closed culture * Compare and contrast the processes of batch and continuous culture * Describe the difference between primary and secondary metabolites * Describe the importance of manipulating the growing conditions in a fermentation vessel in order to maximise the
Free DNA
Transformation Lab Report Introduction Transformation is the transfers of virulence from one cell to another‚ through the transferring of genetic material. It was originally postulated in 1928 through the works of Federick Griffith‚ a British microbiologist. Griffith observed that the mutant form‚ non-virulent form‚ of the bacteria Streptococcus Pnumoniae could be transformed into the normal‚ virulent form‚ when injected into mice along with heat killed normal forms. He concluded that somehow
Premium DNA Gene
added to the isolated gene so as to make the protein present in the sheep’s milk. The second stage was to remove mature eggs from the ovary of a sheep. The isolated AAT gene and promoter were then inserted into a plasmid; this was done by inserting restriction enzymes into the plasmid and they cut the DNA at the same
Premium DNA
purified plasmid for restriction digests‚ and identifying the mislabeled plasmid. The plasmid DNA from a carrier E. coli strain was purified by the boiling lysis. In the boiling lysis method‚ the bacterial cells were given momentary heat treatment in boiling water in presence of lysozyme and triton X-100. Since the plasmid DNA is small in size‚ it comes out from the bacterial cell‚ while the genomic DNA remains trapped in the cell. Successive high-speed centrifugation separated the plasmid DNA from
Premium Molecular biology DNA
fluorescent protein‚ GFP‚ is located near the beta lactamase gene on the pGLO plasmid‚ which protects bacteria from the antibiotic ampicillin. When the cell produced beta lactamase to deactivate ampicillin‚ the GFP gene was also transcribed‚ producing the fluorescent protein observed. In the LB/amp plate containing the +pGLO sample white‚ non florescent cells were observed. While these genes contained the pGLO plasmid and the GFP
Premium Bacteria Gene DNA