Nucleic Acid Flashcards
What are the requirements of gDNA purification?
lyse membrane, denature proteins, remove inactive or endogenous nucleases, precipitate DNA and wash DNA
What are the chelating agents related to endogenous or inactive nucleases, and the process of removing them?
proteinase K - chelating agents
What is the difference between the structure of deoxyribose and ribose genetic material?
Deoxyribose does not include the hydroxyl group in the sugar, whereas ribose does.
Which nucleic acid/s have three hydrogen bonds between the nitrogenous base and the sugar?
Guanine and Cytosine
What chemical group is attached to the 5 primed end of DNA?
Phosphate, bonded to another carbon
What chemical group is attached to the 3 primed end of DNA?
Hydroxyl group
What is the net charge (positive or negative) of the DNA backbone?
Negative
What are some examples of organic solvents used to separate DNA from other genetic material?
Phenol, chloroform
Describe the polarity of organic solvents: chloroform and phenol.
They are both non-polar solvents.
Why does the isolation of genomes method of “salting out” actually work?
The salt infiltrates the solution and gives no room for the proteins to interact with the water molecules and then proceed to precipitate out.
What is the difference between Rna and Dna structure?
Rna has ribose attached to the sugar, whereas Dna has deoxyribose attached to the sugar.
What are some features of RNA that is important to know when isolating RNA?
Ribose sugar not deoxyribose, unstable, single stranded, contains uracil not thymine base and is capable of RNAse degradation.
What does it mean to “lyse” the membrane?
It means to disturb or dissolve the boundary, barrier or lipid natured membrane in order for the nuclease to release genetic material (DNA).
What is spectrophotometric analysis?
Measures quantity and purity of nucleic acid.
How does spectrophotometric quantify and anaylse nucleic acid?
Nucleic acid absorbs UV light.
