Unit 1.2
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What are the basic units of DNA?
Nucleotides
What holds the complementary DNA bases in place?
Weak hydrogen bonds
The order of the bases code for what?
The production of a particular protein
What molecule is the 5’ end?
Phosphate
What molecule is the 3’ end?
Deoxyribose sugar
What end are nucleotides added to?
The 3’ end
What are the requirements for DNA replication?
• DNA template • Primers • Enzymes (DNA polymerase and ligase) • Free nucleotides • ATP
What are stages of DNA replication?
Stage 1: Template strand DNA Stage 2: DNA is unwound and hydrogen bonds between bases are broken to form 2 template strands Stage 3: DNA polymerase adds DNA nucleotides using complementary base pairing, to the deoxyribose (3’) end of the new DNA strand which is forming. Stage 4: The lagging strand is replicated in fragments Stage 5: Fragments of DNA are joined together by ligase Stage 6: DNA strand winds back up to form 2 identical double helixes
What is Polymerase Chain Reaction (PCR)?
A technique for the amplification of DNA in vitro.
The amplification of DNA uses the complementary of what?
Primers
The complementary primers are short strands of what?
Nucleotides
Why are 2 separate primers needed for PCR?
There are 2 different strands
What are the requirements for PCR?
• Primers • Free nucleotides • Sample DNA • Heat resistant DNA polymerase
Stages of PCR?
Stage 1: DNA is heated to between 92-98 degrees celsius to break the hydrogen bonds and separate the strands Stage 2: DNA is cooled to between 50-65 degrees celsius to allow primers to bind to their target sequence Stage 3: DNA is heated again to between 70-80 degrees celsius to allow heat-tolerant DNA polymerase to replicate the region of DNA
What is particular about the polymerase enzyme used in PCR?
So that the heat-tolerant polymerase don’t denature