DNA Replication Flashcards
What does PCR stand for and what does it mean?
Polymerase Chain Reaction, = synthesis of
DNA by thermostable
DNA polymerase in the
test tube
Define DNA Cloning
Prior to the 1980’s, the primary method for producing many copies of a gene
What is the key element of PCR?
Heat. DNA is submitting through heating and cooling cycles.
Explain the steps of PCR and how it works
Denaturation, Annealing, and Primer Extension. GATHER, BOUND, POLYMERASE, TO MIMIC THE PROCESS OF DNA REPLICATION First, a small amount of DNA is gathered, a pair of primers are used to bind each end of the target sequence, Then a dna polymerase is used, then Four dNTPs (i.e., dATP, dCTP, dGTP, dTTP), and last a few essential ions and salts. They use those ingredients to to mimic the natural dna replication process. A thermocycler is used with cool and heat.
What is a Thermocycler?
A machine that jump-starts each stage of the reaction by raising and lowering the temperature of the chemical components at specific times and for a preset number of cycles.
What Denatursation?
The melting of DNA into individual strands. The started solution is heated to break the bonds joining the strands of a DNA double helix enabling it to separate into two strands.
What is Annealing?
The reaction mixture is quickly cooled, usually between 30 and 65 degrees. This gives the primers an opportunity to bind or anneal to their complementary sequences on the single strands of DNA
What is Extension?
The sample is heated and the DNA polymerase begins to make a new DNA strand by attaching to the primers
What is (RT-PCR)?
Reverse Transcription Polymerase Chain Reaction, this combines real time PCR and reverse transcription
What is Reverse Transcription?
The process that makes dna from rna
What is qPCR?
quantitative real-time PCR (qPCR)
What is PCR used for?
Sequencing, cloning, and analysis in the case of forensics
What are the two primers called?
Forward and Reverse primers
What can we tell if both primers attach to the same piece of DNA and synthesize in the same direction?
That there is no PCR product
Explain the process of replication
Template strands separate, New nucleotides are added according to the templates, and they are joined into the new strand
T/F Replication occurs prior to cell division
True
T/F Replication happens during the G1 phase of the cell cycle
False. It happens during the S phase.
T/F The cell divides producing
two daughter cells during the
mitotic phase
True