PCR Flashcards
What is the purpose of P.C.R?
Allows a quantity of DNA to be amplifyied for analysis.
What does P.C.R stand for?
Polymerase Chain Reaction
What is the first stage of P.C.R and what happens?
Denaturation: The DNA sample is heated to 95 degrees to separate the double stranded DNA into two single strands
What is the second stage of P.C.R and what happens?
This stage is know as Annealing. This cools down the strand 50-60 degrees allowing primers (6-26 Bp long) to bind to complementary sequences of each DNA strand.
What is a primer?
Short sequence of DNA that flanks the target region
What is the third stage of P.C.R?
Extension: The sample is heated to 70 degrees. Thermally Stable Polymarase (TAQ) are added, which increase length of DNA to 56Bps long by making bonds in the sugar phosphate.
What is the final stage of P.C.R?
Repeat: The process is repeated 40 times, resulting in an exponential increase in the target sequence.
Name the stages of P.C.R in order
- Denaturation
- Annealing
- Extension
- Repeat