1. Polymerase Chain Reaction
PCR
Polymerase Chain Reaction

2. What is PCR?
PCR intro:

3. PCR requires the following-

4. Reaction Components
Template DNA -

5. Reaction Components DNA primers In the cell (in vivo), primers are
that serve as a starting point for DNA replication
In PCR (in vitro), Primers are
that exactly match the beginning and the end of the DNA fragment to be amplified
Reverse Primer
Forward Primer
Target DNA length 3’ 5’
Genomic DNA

6. Reaction Components
DNA polymerase -

7. Reaction Components
DNA Polymerase
Why?

8. Reaction Components
dNTPs - A T G C

9. Reaction Components Buffer
To work properly, Taq polymerase needs magnesium ions:
* Too little:
* Too much:

10. A three- step process
Denaturation:
Annealing:
Extension:

11. How Does PCR Work? Step 1: Denaturation
Double Stranded DNA target
Denatured single strand
Denatured single strand

12. How Does PCR Work? Step 2: Annealing - Primer Binding to Target also called ________________5’ 3’ 5”
Template Strand #1
Reverse Primer
Template Strand #2
Forward Primer

13. How Does PCR Work? Step 3: Extension5’ 3’
Taq
Template Strand #1
Template Strand #2

14. Equipment Thermal cycler
Thermal cyclers have metal heat blocks with holes where PCR reaction tubes can be inserted. The thermalcycler then

15. PCR a CHAIN reaction
At the end of the first PCR cycle, there are now

16. Exponential Amplification of the Target DNA Sequence

17. http://highered. mheducation

18. BEWARE ! Sources: VALIDATION
Do a_______________________________to validate that the PCR product is amplified from the intended DNA, not some other source of DNA.
________________________________________________validates that the reaction conditions and thermal cycler work properly.

19. PCR: Analysis
At the end of a PCR reaction, there is a A LOT more of your target DNA than before the reaction started…billions of copies!
Now the sample is large enough
PCR can identify
PCR can

20. PCR A powerful, versatile tool
There are many uses for PCR in a endless array of scientific fields: