Molecular Biology Lab Help?

Posted on 2014-04-15 08:24:00 in Biology by LAWRENCE

Here are a few questions I have that I did not get full credit on in homework or test questions. Any answer you are able to give, whether to one question, a couple, or all of them, would be incredibly helpful!



1. Compare (3 things) and contrast (3 things) PCR and probe labeling.



2. If you isolate the following mRNA from S. cerevisiae, what would you expect the coding (or sense strand) DNA sequence it came from to be?

mRNA: 5'AUG CGU UGC CCG UUA UAG3' (I thought I did this correctly, but apparently something was wrong and I'm not sure what)



3. What does reverse transcriptase do? Name one place it is found in nature.



4. Why can you not design RT-PCR primers against an intron?



5. How could you design an RT-PCR primer that would be unlikely to give a false positive reading from DNA contamination?



6. What are two ways to make a cell "competent"? Why do we make competent cells?



7. T4 ligase buffer contains the following--50mM Tris-HCl, 10mM MgCl2, 1mM ATP, 10mM DTT--explain a likely purpose of these components in the buffer.



8. What problem would we encounter if our DNA linkers were degraded by a single base pair?



9. In plasmid purification protocol, what would happen if you forgot to incubate E.coli with TSE (25mM Tris, 0.3M sucrose, 2.5mM EDTA)?



10. In plasmid purification protocol, what would happen if you forgot to incubate E.coli with 7.5M ammonium acetate?

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