DNA Manipulation and Techniques Flashcards
What does PCR stand for?
Polymerase Chain Reaction
How would scientists increase the amount of a sample of DNA?
By using PCR
What is the function of Polymerase Chain Reaction
It amplifies and makes more copies of DNA samples
What are the three stages of polymerase chain reaction?
Denaturation, Annealing, Elongation
What materials are required for polymerase chain reaction?
DNA strands, 2 primers, buffer, Tac polymerase
Why is Tac polymerase used instead of DNA polymerase?
Because Tac polymerase has a higher temperature for its optimum range. It can withstand the high temperatures during PCR.
What would happen if DNA polymerase was used in polymerase chain reaction?
DNA polymerase would denature as it cannot withstand such high temperatures. It would denature and essentially die.
What happens in the denaturation stage?
The DNA strand is heated to 90-95 degrees C to break the hydrogen bonds between the strands to seperate them.
What happens in the annealing stage?
The DNA strands are cooled to 50-55 degrees C to allow for primers to bind to the strands.
What happens in the elongation stage?
The DNA strands are reheated to 72 degree C to allow Tac polymerase to bind to the primers and begin synthesising identical DNA strands in a 5’ to 3’ direction.
What happens at the end of the PCR?
The process repeats in order to increase the amount of the sample.
What is gel electrophoresis used for?
Gel electrophoresis is used to seperate DNA fragments and measure their length.
