POLYMERASE CHAIN REACTION DNA SEQUENCING TECHNOLOGIES Flashcards
POLYMERASECHAIN REACTION developed by american biochemist _____ in the
_______s.
Kary Mullis in the1980s.
a vital technique in molecular biology, enabling
researchers to amplify specific DNA fragments
exponentially. This is a powerful lab technique used to generate billions of copies of a specific sequence of DNA.
POLYMERASE CHAIN REACTION (PCR)
What are the NECESSARY MATERIALS for PCR?
DNA Template
Special Polymerase (Taq Polymerase)
dNTPs
Primers
All PCR reagents are combined in the PCR tube in a buffer that ensures most PCR product. _____ and _____ ion concentrations are particularly important and sometimes need to be adjusted.
Magnesium and Potassium ion concentration
PCR Requires:
1.
2.
3.
4.
5.
- Buffer
- Template DNA
- Primers
- dNTPs
- Taq polymerase
In this stage, DNA is heated to a
minimum 95 °C The heat breaks the hydrogen bonds of DNA template and separates into single strands.
- Denaturation
3 stages of PCR cycle
- Denaturation
- Annealing
3.Elongation
In this stage, DNA is then cooled
down to 45-70°C the
the DNA primers bind to
the individual single strands.
- Annealing
In this stage, DNA polymerase insert nucleotides and extend the newly strand.
Elongation: 3rd stage
During ____ stage, the temperature is
raised to 72°C. The polymerase uses
the annealed primers as the starting
point for DNA synthesis.
- Elongation
PCR 1st cycle produce how many copies?
4 copies
2nd cycle of PCR produce?
8 copies
3rd cycle of PCR produce?
16 copies
3th cycle of PCR produce?
32 copies
is the process of
determining the nucleic acid
sequence – the order of nucleotides
in DNA.
DNA sequencing
