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- biotechnology lab class :1. The bacterial streaking on antibiotic containing agar plates has been completed and the plates will be available for you to inspect in the lab class.2. You will complete a restriction enzyme digest on the four available plasmid stocks and then examine the products by agarose gel electrophoresis. Photographs of the gel are attached to identify the plasmid present in each analysed sample.Now consider the following questions :- For each of the four plasmids used in the practical, identify the size (in kb) of the linear DNA fragment(s) that you would expect to obtain if the EcoRI digest is complete.- Using the provided negative image of the gel, which shows the bands present in the ND and D samples for each plasmid, identify the plasmid that was present in each of the four plasmid stock.- Explain how you were able to identify plasmid in each sample using the gel, considering how linear DNA fragments of different length are separated by agarose gel…DNA Cheek Cells Lab Identify the component of the solution which is the extracted DNA10. When looking at your agar plate, where are the bacteria that did not take up the plasmid during the transformation? (It is a 4 letter word). Thanks for the help!
- 5- What’s the purpose of the DNA extraction? a- Force the cell to make more DNAb- Kill the cells.c- Lyse the cells in a way that the DNA get releasedd- Make the cells release DNA in vesicles 6- What amount of agar do you need to weigh if you want to make 300mL of a 2% (w/v)agarose gel?n gel electrophoresis of DNA, the different bands in the final gel form because the DNA molecules _______. a. are from different organisms b. have different lengths c. have different nucleotide compositions d. have different genesDrag and drop the appropriate text in the gaps below. Firstly, the plasmid DNA in the E. coli is propagated through Then the bacterial cells are pelleted by centrifugation at 10,000 RPM. The media supernatant is removed and the bacterial cells are The bacterial cells are then lysed via |. Contaminating macromolecules such as protein and chromosomal DNA is removed by Overnight incubation of the bacterial culture washed in cell resuspension buffer addition of lysis buffer addition of neutralisation buffer washing of the spin column separation via agarose gel electrophoresis
- 2. Restriction Enzyme Mapping - use any resources to assist you including the hints below. A circular plasmid molecule (12,000 total base pairs in size) was cut with a series of restriction enzymes and the digestions were size fractionated by agarose gel electrophoresis. Some digestions involved just one enzyme (single digest), some combinations of two enzymes (double digests), and one utilized all three enzymes (triple digest). Agarose gel electrophoresis of the digestions produced bands of the following sizes. Enzyme EcoRI Hindi!! Pstl EcoRI and Hindill EcoRI and Psti Hindill and Pstl EcoRI, Hindill, and Pst I Bands of the Agarose Gel (size in base pairs) 2300 and 9700 4000 and 8000 12,000 800, 1500, 2500, and 7200 2300, 3200, and 6500 4000 800, 1500, 2500, 3200, and 4000 I Draw a plasmid map showing the location of the restriction enzyme sites relative to each other for each map. Include all 7 maps in your answer.What is the correct order for the steps of transformation given inthe following list?1. Recombination with the bacterial chromosome2. Binding of a large DNA fragment to the surface of a bacterialcell3. Cutting a large DNA fragment into smaller pieces4. Uptake of DNA into the cytoplasm5. Degradation of one of the DNA strandsa. 1, 2, 3, 4, 5b. 2, 3, 5, 4, 1c. 2, 3, 4, 5, 1d. 2, 5, 4, 3, 1Some Steps Involved in Creating Recombinant DNA 1. Plasmid DNA is cut at the restriction site.2. Foreign DNA is cut at the restriction site.3. Plasmid DNA is opened and sticky ends are formed.4. Foreign DNA restriction fragments are isolated5. Target sequence in the plasmid is recognized by the restriction endonuclease.6. Target sequences in the foreign DNA are recognized by the restriction endonuclease.7. Foreign DNA restriction fragment is inserted into the plasmid at the restriction site.8. DNA ligase splices the foreign restriction fragment and the plasmid together. Identify the correct sequence of steps involved in cutting and reassembling DNA molecules to make recombinant DNA. Start with preparation of the plasmid DNA. Select one: a. 6, 2, 4, 7, 8, 5, 1, 3 b. 5, 1, 3, 6, 2, 4, 8, 7 c. 5, 1, 3, 6, 2, 4, 7, 8 d. 6, 2, 4, 5, 1, 3, 8, 7
- Part A. If student counts 63 colonies on their 10^-6 dilution LB plate. What was the original concentration of their cells if they plated 100ul? Part B.If we used pGFPuv as the template for PCR positive control. This is because: a. it contains the GFP gene so it should show a product. b. It contains DNA fragments that were added to the ligation reaction. c. It is the desired plasmid we wanted to make. d. So we have a band to compare our unknown plasid to allowing us to check if the unknown is the right size.Virology 8. A positive sense RNA sequence is given below. Give the correct sequence of the CDNA first strand synthesised by reverse transcriptase. 5'-UUCCGAUAUCGGCACUGAGUA-3' 5'- -3'The figure below illustrates the stages of production of useful medical products using recombinant DNA technology. Closely study the figure then answer the Activity 1 questions that follow: Human cell Bacterium 1. DNA Plasmid DNA Human insulin-producing gone 2. DNA is cut with restriction Bacterial DNA with human gone inserted enzymes. 3. Plasmid is reintroduced into bacterium. 4. Engineered bacteria multiply producing insulin. 5. Insulin is separated and purified to produce human insulin. Human insulin Itulin 6. Insulin injected into patient 1. What is a plasmid? Make a diagram of bacterial cell containing a plasmid. 2. How do you explain the selection of plasmids for carrying the desired gener 3. Follow the steps of human insulin production, then 4. Make a diagram for the production of growth hormone. first