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Genetics Q4
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- Question one: PCR You want to amplify the underlined sequence, 1) Design Forward and Reverse primers to do so 2) Calculate the melting temperature for them 3) Calculate annealing temperature for them 4) Do you think your primers are high quality (Length, differences in melting between R and F, possibility of forming hairpins)? GTGTGCTGGTTATTCAAACAGATAAAAAAATTAAT CTATATGGTAATGCTCTAAGCCGCGCAAATACAG AATATGTGCCAGCCTCTACATTTAAAATGTTGAAT GCCCTGATCGGATTGGAGAACCAGAAAACGGATA TTAATGAAATATTTAAATGGAAGGGCGAGAAAAG GTCATTTACCGCTTGGGAAAAAGACATGAСАСТА GGAGAAGCCATGAAGCTTTCTGCAGTOCCCAGTCT ATCAGGAACTTGCGCGACGTATCGGTCTTGATCT CATGCAAAAAGAAGTAAAACGTATTGGTTTCGGTA ATGCTGAAATTGGACAGCAGGTTGATAATTTCTG GTTGGTAGGACCATTAAAGGTTACGCCTATTCAA GAGGTAGAGTTTGTTTCCCAATTAGCACATACACA GCTTCCATTTAGTGAAAAAGTGCAGGCTAATGTAA AAAATATGCTTCTTTTAGAAGAGAGTAATGGCTAC AAAATTTTTGGAAAGACTGGTTGGGCAATGGATAT AAAACCACAAGTGGGCTGGTTGACCGGCTGGGTT GAGQuestion 6 Plasmids that generate blunt ends after restriction enzyme digestion are the ones most effective for recombinant DNA technology. A True B) FalseQuestion 43 The addition of restriction endonucleases in the cloning process is done following the ligation with DNA ligase. A) True B) False
- Question 3 PUC plasmids are widely used because of the beta-lactamase gene that renders an easier blue and white screening process in transformant selection. (A) True B) FalseQuestion 3Which of the following statements regarding conjugation pili is correct? Question 3 options: a) They facilitate the transfer of RNA between bacterial cells b) They contribute to antibiotic resistance c) They are involved in sexual reproduction d) They are longer than fimbriae and flagellaQUESTION 1 You want to perform PCR on the CDNA of the spike gene from a SARS CoV-2 sample so that you can sequence it. Based on the sequence below, which of the following primer pairs would probably work for PCR of this gene? Spike gene Sequence: 5' ATGTTTATTTTCTTATTATTTTTTACTCTCACTAGTGGTAGTGACCTTGACCGGTGCACCACTTTTGATG ATGTTCAAGCTCCTAATTACACTCAACATACTTCATCTATGAGGGGGGTT TACTATCCTGATGAAATTTT. .. (it's really long so didn't post the whole thing.).TCTTGCTTTGTTGCATGACTAGTTGTTGCAGTTGCCTCAAGGGTGCATGCTCTTGTGGTTCTTGCTGCAA GTTTG ATGAGGATGACTCTGAGCCAGTTCTCAAGGGTGTCAAATTACATTACACATAA 3' Forward primer: 5' - CTC TCA CTA GTG GTA GTG ACC - 3' (Tm = 60.5 °C in a standard qPCR mix) Reverse Primer: 5' GGG TGT CAA ATT ACA TTA CAC ATA - 3' (Tm= 59.6 °C in a standard QPCR mix) Forward Primer: 5'- ATG TTT ATT TTC TTA TTA TT -3' (Tm=D 47.2 °C in a standard qPCR mix) Reverse Primer: 5'- GCA AGA ACC ACA AGA GCA TGC ACC -3' (Tm= 68 °C in a standard qPCR mix) Forward primer: 5' - CTC TCA CTA GTG GTA GTG ACC -3'…
- Question 14 If Guanine is 30% of the total bases in a dsDNA, the thymine content is 40%. A) True B) FalseQuestion 49 The Sanger method of DNA sequencing follows the principle of complementarity just like in the replication process. A) True B) FalseQuestion 2 The discovery of the DNA structure started the modern era of biotechnology. A) True B False
- Question #3: CRISPR has been used to cure an individual from sickle cell. Below is a Sanger electropherogram of a sequence from a patient without sickle cell and one with sickle cell. Sequence from a normal individual mmmm Sequence from the diseased individual G T GIIC A GC A Se SCIENCEphe A G A SCIENCE SCIENCEphoto G a) Where is the change in the sequence and what is the consequence to the protein sequence of this mutation? b) Below is an image of the normal and diseased quaternary hemoglobin protein. What is different about the protein shape and why does that structure have a huge impact on its function (please name the function!)? Adult haemogBRAR G G G G A G Sickle Cell haemoglobin S Structure a s RARY COLIBRARY c) If you were to use CRISPR to modify the genome of a diseased individual, to which nucleotides might you design your guide RNA? Why? d) RNA Seq is used to determine off-target effects of Cas9 cleavage. Why is this an appropriate tool to determine these effects? e) Data on…Recombinant pharmaceuticals (for the production of insulin, human growth hormone or blood clotting factors) Question: What are controversies or ethical dilemmas surrounding this genetic technology or process?Question 13 If a recombinant plasmid (below) was obtained inserting DNA into the BamHI site, screening for the recombinant plasmid can be done by which of the following technique? A) Plate on agar plates containing tetracycline and ampicillin. (B) Plate on agar plates containing ampicillin, C) Plate the cells on agar containing ampicillin then surviving colonies are plated on another agar plates with tetracycline. D) Plate on agar plates containing tetracycline. Pal Pord- ampr EcoRI ПРИ Ano pBR322 (4363 bases) Prull BamHI Sall let" Aval -Sall