Erstellt von Sophia Khan
vor fast 9 Jahre
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Frage | Antworten |
What was the goal of lab 3? | ligate the DNA fragments produced during Lab 2, using DNA ligase to make new recombinant plasmids (pARA-R) |
What was the goal plasmid to make during this lab? Indicate the important features of this particular plasmid and the function of each feature. | pARA-R - ampicillin resistance (ampR): allows for identification of bacteria that have taken in the plasmid - rfp gene - promoter sequence (pBAD): initiating transcription - arabinose activator sequence (araC): - ori sequence: initiating DNA replication |
What is the role of DNA Ligase in replication? What about in gene cloning? | - replication: one of the daughter strands is made up of DNA fragments and the ligase joins the fragments by catalyzing the formation of covalent bonds between adjacent nucleotides - gene cloning: the ligase catalyzes bonding along DNA's sugar-phosphate backbone after the bases in complementary sticky ends have formed hydrogen bonds to each other |
Why did we need to inactivate BamHI and HindIII before beginning this lab? | otherwise it would cut/denature the DNA, when the goal of the lab is to ligate the fragments |
What was in the LIG tube? What happened in this tube? | pKAN-R fragments, pARA-R fragments, buffer, and water |
What properties of the DNA restriction fragments allow for ligation to occur? | complementary sticky ends; the ligase catalyzes bonding of the backbone only after the base pairs have been created |
Could two rfp-pBAD fragments form during ligation? Why or why not? | Yes, the two fragments can join because they have complementary sticky ends |
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