DNA Renaturation Flashcards
Define annealing
The reuniting of DNA strands
What is another term for renaturation?
Annealing
What three factors contribute to renaturation efficiency?
Temperature
DNA concentration
Time
What temperature is best for annealing?
Tm - 25C
Why is Tm - 25C ideal?
It is low enough that it doesn’t promote melting but high enough to allow for rapid diffusional movement of DNA and weakens transient bonding between sequence mismatches
What prevents annealing? What is the term?
Rapid cooling of denatured DNA
Quenching
What are the two ways the term ‘quenching’ can be used?
1) when nitrogenous bases are close together
2) rapidly cooling a DNA sample to keep it denatured
What is the effect of DNA concentration on annealing?
The Higher DNA concentration, the faster the annealing
Why does a high DNA concentration increase the rate of annealing?
Has a greater likelihood of its complementary strands encountering each other within a given amount of time
How does time affect renaturation?
The longer the time allowed for annealing, the more renaturation will occur
What is the effect of DNA complexity?
Contributes to the efficiency of DNA renaturation
What is a fourth factor in renaturation?
DNA complexity
How does DNA complexity contribute to efficiency?
The more complex the DNA, the slower or less efficient the renaturation
