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True/False (10 Questions • 20 Points) Instructions Read each…

True/False (10 Questions • 20 Points) Instructions Read each statement carefully. Decide whether the statement is True or False based on your understanding of the course material. Remember that if a statement is 99% true, it is 100% FALSE Each question is worth 2 points. There is only one correct answer.

NOTE: This question (Parts I to IV below) will be graded sep…

NOTE: This question (Parts I to IV below) will be graded separately and it is worth 20 points. Please complete the following questions on a blank piece of paper. You will have 30 minutes after you finish your test to submit all the answers for this question to the “Exam 3 Part 2” assignment on the “Exams” link. Please submit the file as a pdf file. The questions will only be shown in the actual exam, so it is important to collect all the data/information before submitting/disconnecting from your exam.  Show both sides of your paper (scratch portion and essay) to the camera before submitting and exiting the Exam. You have been recruited by a covert biotech startup to help mass-produce the protein coded by the “ParalyzX1” gene produced by a mutualistic bacterium, which gives a bioluminescent deep-sea eel its rapid neuromuscular paralytic toxin. A field operative has provided you with clean, uncontaminated DNA. The sequence of the gene, isolated from the bacteria that inhabit the eel’s venom gland, is shown below: 1 11 21 31 41 CATCGGTACG  TAGCCTAGGA  GCAATTGACA CGATCGGATT ACGTAGCTAG 51 61 71 81 91 CATGTATAAT GCGTACGCTA CCGGATCTAG AAGGAGGTAT  ATGCTGAGCG 101 111 121 131 141 TTGATGGACG TAAATTCGCA TGGTGTTATG AAAATATTCC ACGAACCAGC 151 161 171 181 191 AGGTTGGTGC  ACTAAGCTAG CGGGGGGGAA  ACCCCCCCCT  TTTTTTTTTC 201 211 CGGATCCGTA C

Part II.  Building the recombinant DNA construct. You are r…

Part II.  Building the recombinant DNA construct. You are ready to follow the major steps required to create a recombinant DNA (your gene of interest inside a vector), from amplification of the gene to verifying that your end product doesn’t have an unexpected mutation. A. For the following three steps, name the key enzyme needed to complete each task. (3 pts) 1. PCR and Gel electrophoresis – Key enzyme:___________ 2. Cleaving DNA –  Key enzyme:___________ 3. Ligation into a vector that has the Amp resistance gene – Key enzyme: _____________   The next step will be transformation. For this step, you will use LB Amp plates with X-Gal. B. Assuming your transformation efficiency with the recombinant plasmid was good, but not perfect, how will that plate look? Draw AND explain your result in one sentence. (2 pts)   C. Your research mentee, distracted while checking a group chat, forgot to add the recombinant plasmid to the transformation mix. How would his plate look after the transformation attempt? Draw AND explain your result in one sentence. (2 pts)   Part III. Verifying the PCR product by gel electrophoresis. You performed a PCR to amplify the open reading frame (from the start codon to the stop codon) of the ParalyzX1 gene, plus an extra 10 bp on each end containing the restriction sites used for cloning. You want to run a gel to check whether the PCR product is the expected size. A. Draw the gel electrophoresis results, assuming the PCR worked perfectly. Include a DNA ladder (with 50 bp, 100 bp, 150 bp, and 200 bp bands), positive control, ParalyzX1 sample, and a negative control. Label each lane accordingly. (4 points). B. Also indicate the expected size of the amplified fragment. (0.5 points)