10 Questions
What is the primary purpose of transferring DNA onto a membrane in Southern blotting?
To determine the presence of a specific DNA sequence within a large, complex DNA sample
What is the role of probe in Southern blotting?
To identify which DNA fragments obtained from a digest of a larger DNA clone/genome that contains sequence complementary to a specific probe
What determines the amount of DNA required for restriction enzyme digestion in Southern blotting?
Type of DNA sample (e.g., plasmid DNA, genomic DNA)
What is the purpose of DNA fragmentation using restriction endonucleases in Southern blotting?
To obtain DNA fragments for further analysis
Who invented Southern analysis, a key technique in DNA recombinant technology?
Edwin M. Southern, an English Molecular Biologist
What is the purpose of using purine bases instead of pyrimidines during DNA treatment with diluted acid?
Purine bases are less likely to be depurinated compared to pyrimidines.
Why is capillary action used for the transfer of DNA onto the membrane?
Capillary action allows for the rapid transfer of DNA fragments.
What is the role of 0.4 M NaOH in the transfer of DNA onto a nylon membrane?
It enhances the binding of DNA onto the nylon membrane.
Why is an alkaline buffer used for transferring DNA onto a nylon membrane?
To enhance the binding of DNA onto the nylon membrane.
What is the main reason for using 20X SSC for transferring DNA onto a nitrocellulose membrane?
To improve the stability and binding of DNA onto the nitrocellulose membrane.
This quiz covers the basic concepts of Southern blotting, including the method, lab report questions, and the need for transferring DNA onto a membrane. It also includes the process of DNA transfer, solid phase membrane support, and probe hybridization.
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