18 Questions
What was the purpose of digesting the nucleic acids with RNAase A?
To remove RNA from the DNA preparation
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
What was the composition of the DNA preparation used in these experiments?
1/3 supercoiled plasmids, 1/3 nicked circles, and 1/3 larger oligomers of the plasmid
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
What was the volume of DNA solution injected into the male pronucleus?
Approximately 2 pl
What was the purpose of washing the ova free of cytochalasin B after the injection?
To allow the ova to resume normal development
What was the purpose of the thymidine kinase assay described in the text?
To measure the activity of thymidine kinase in mouse tissues
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
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
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
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
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
What was used to label DNA probes in the described experiment?
32P-dNTPs
Which enzyme was NOT mentioned in the text for restriction digestion?
Sal I
What was the purpose of using RNAase A in the restriction digests?
To degrade RNA contaminants
What specific conditions were used for the hybridizations in the described experiment?
50% formamide, 3.2 × SSC, 10% dextran sulfate
Which fragment was used to prepare the 32P-cDNA?
460 bp fragment from Bgl II site to Sst I site
What method was used to transfer DNA onto nitrocellulose in the experiment?
Southern blot hybridization
Test your knowledge on the digestion of nucleic acids with RNAase A and separation through Bio-Gel A50m column. Learn about the different plasmid structures revealed by agarose gel electrophoresis. This quiz also covers the injection and manipulation of eggs.
Make Your Own Quizzes and Flashcards
Convert your notes into interactive study material.
Get started for free
