BISC 440 Lab 5: Molecular Cloning questions and answers 100% accurate

BISC 440 Lab 5: Molecular Cloning questions and answers 100% accurate The RAD52 gene can be isolated from the yeast cells. What should we do first so we can isolate the gene we need? a. separate the DNA using gel electrophoresis b. perform DNA sequencing c. use a restriction enzyme d. perform DNA extraction d In order to isolate RAD52, what should we do after extracting all DNA from the yeast cells? a. sequence the DNA b. proceed to the ligation step c. transform the yeast cells d. amplify the RAD52 gene d How can we cut the eGFP gene from the plasmid? a. isolate the DNA using phenol-chloroform extraction b. perform restriction enzyme digestion c. grow up the plasmid in bacteria so they secrete the gene d. perform DNA sequencing on the whole plasmid b How can you confirm that we have successfully extracted the DNA from the yeast? a. weigh it using an analytical scale b. analyze it using gel electrophoresis and an analytical scale c. analyze it using the UV transilluminator d. analyze it using gel electrophoresis and the NanoDrop d What can you conclude about the size of the RAD52 gene? a. the RAD52 gene is about 3500 bp long b. the RAD52 gene is precisely 1416 bp long c. the RAD52 gene is about 1500 bp long d. the RAD52 gene is precisely 2543 bp long c Where is the eGFP gene located? a. at position 1-267 b. at position c. at position 702 d. at position d Which two restriction enzymes cut this plasmid once and can be sued to isolate the eGFP gene? Find the single restriction sites a. Notl and EcoRI b. Xbal and Xhol c. Ncol and Xhol d. we only need to use one enzyme b In order to ensure that the restriction enzymes work properly, we need to select the optimal buffer and incubation temperature. Which buffer should we choose for cutting the eGFP using Xbal and Xhol? a. buffer 3 b. buffer 2 c. buffer 1 d. buffer 4 a Which incubation temperature is optimal for cutting eGFP from its plasmid? a. 25°C b. 65°C c. 37°C d. 80°C c Which of the following restriction enzymes produces blunt-end DNA strands? a. Xhol b. EcoRI c. HindIII d. EcoRV d What is the next step after performing gel electrophoresis to obtain the pure eGFP fragment that has been cut from its plasmid? a. perform enzymatic DNA ligation b. perform restriction enzyme digestion c. perform an electrophoretic gel extraction d. perform spectrophotometric NanoDrop analysis c eGFP was isolated from the plasmid pPyCAG-eGFP-IP using Xbal and Xhol restriction enzymes. Based on this information, which band contains the eGFP? a. A b. D c. B d. C d Which of these DNA orientations is correct when ligating RAD52 and eGFP? a. figure C b. figure D c. figure A d. figure B c Which of the following techniques do you use in order to insert exogenous DNA into the yeast calls? a. transformation b. transduction c. all of the options d. conjugation a We know that the exogenous DNA cannot enter the yeast cell by itself. What should we do to make the transformation successful? a. make the growth medium richer b. create competent yeast cells c. amplify the exogenous DNA d. add antibiotic to the medium b It is common that only some of the yeast cells will successfully take up the plasmid. How do we determine which yeast contain the pTRE-RAD52-eGFP plasmid? a. through gel electrophoresis b. it cannot be determined c. by sequencing each colony d. through antibiotic selection d Which colony should we pick for the expression analysis? a. C b. A c. B d. D b