Nucleic Acids Digestion and Separation Quiz

Play an AI-generated podcast conversation about this lesson

What was the purpose of digesting the nucleic acids with RNAase A?

To separate DNA from RNA fragments

To remove RNA from the DNA preparation (correct)

To ensure the DNA was in a supercoiled form

To degrade any remaining RNA in the sample

What was the purpose of passing the digested nucleic acids through the Bio-Gel A50m column?

To further purify the DNA from the RNA fragments (correct)

To concentrate the DNA for downstream applications

To remove any residual RNAase A from the sample

To separate the plasmids based on their size

What was the composition of the DNA preparation used in these experiments?

Mostly supercoiled plasmids with a small amount of nicked circles and linear fragments

1/3 supercoiled plasmids, 1/3 nicked circles, and 1/3 larger oligomers of the plasmid (correct)

1/2 supercoiled plasmids and 1/2 nicked circles

1/3 supercoiled plasmids, 1/3 nicked circles, and 1/3 linear DNA fragments

What was the purpose of using cytochalasin B during the injection of the DNA into the pronucleus?

To increase the viscosity of the medium to stabilize the egg during injection Signup and view all the answers

What was the volume of DNA solution injected into the male pronucleus?

Approximately 2 pl Signup and view all the answers

What was the purpose of washing the ova free of cytochalasin B after the injection?

To allow the ova to resume normal development Signup and view all the answers

What was the purpose of the thymidine kinase assay described in the text?

To measure the activity of thymidine kinase in mouse tissues Signup and view all the answers

What was the main purpose of the nucleic acid isolation procedure described in the text?

To extract and purify DNA and RNA from mouse tissues Signup and view all the answers

What was the purpose of the $eta$-mercaptoethanol included in the thymidine kinase assay reaction mixture?

To act as a reducing agent and maintain the activity of thymidine kinase Signup and view all the answers

What was the purpose of the proteinase K used during the nucleic acid isolation procedure?

To degrade proteins and facilitate the extraction of nucleic acids Signup and view all the answers

What was the purpose of the DE-81 paper used in the thymidine kinase assay?

To adsorb the $^3$H-TMP produced during the enzyme assay Signup and view all the answers

What was the purpose of the NaF included in the thymidine kinase assay reaction mixture?

To inhibit the activity of phosphatases and prevent the dephosphorylation of $^3$H-TMP Signup and view all the answers

What was used to label DNA probes in the described experiment?

32P-dNTPs Signup and view all the answers

Which enzyme was NOT mentioned in the text for restriction digestion?

Sal I Signup and view all the answers

What was the purpose of using RNAase A in the restriction digests?

To degrade RNA contaminants Signup and view all the answers

What specific conditions were used for the hybridizations in the described experiment?

50% formamide, 3.2 × SSC, 10% dextran sulfate Signup and view all the answers

Which fragment was used to prepare the 32P-cDNA?

460 bp fragment from Bgl II site to Sst I site Signup and view all the answers

What method was used to transfer DNA onto nitrocellulose in the experiment?

Southern blot hybridization Signup and view all the answers