1. PCR Related Products

2. Current Products Thermostable DNA polymerases TaqExtender
Pfu
PfuTurbo
TaqPlus Precision
TaqPlus Long
Taq
TaqExtender
Perfect Match
Opti-Prime

3. Components of PCR Thermostable DNA polymerase Reaction Buffer Primers
Template DNA
dNTPs
Water
Cycling Conditions
Each component affects the quality of the PCR

4. PCR Qualities Yield Fidelity Specificity Length
These are the qualities that we can measure and optimize.
Usually one quality has to be sacrificed to improve another.

5. Thermostable DNA polymerases
Pfu
PfuTurbo
TaqPlus Precision
TaqPlus Long
Taq

6. error rate extension Recommended PCR Target (x10 ) time Length (kb)-6 )
time
Length
(kb)
required
(min/kb
vector
genomic
target)
targets
targets
PfuTurbo DNA
1.3 +
0.2 1
0-10
0-6
polymerase 2
0-15
0-10
Pfu DNA
1.3 +
0.2 2
0-10
0-4
polymerase
TaqPlus
3.9 +
1.3 1
0-15
0-10
Precision
system
TaqPlus Long
7.6 +
1.2
0.5-1
0-35
0-18.5
system
Taq DNA
8.0 +
3.9 1
0-10
0-5
polymerase

7. Functional Domains of Pfu DNA Polymerase
5’-3’
exo
DNA-dependent DNA
polymerase
3’-5’ exo
non-
functional
proofreading DNA synthesis
activity

8. Fidelity Assay PCR Mutations are detected in the lacI gene
not single round extension
Mutations are detected in the lacI gene
Color detection from Big Blue system
Results have been published in peer reviewed journals

9. Characteristics of Pfu
High Fidelity
Native Hot Start activity
High Thermostability
Slow polymerization rate
Available as native and cloned enzyme

10. Accuracy of Polymerases in PCR

11. Extension Time Pfu requires 2 min/kb
Minutes of extension time/kb of PCR target M
Taq
Pfu

12. Activity vs Temperature Pfu and Taq
percent activity
Incubation Temperature (degrees C)

13. Pfu: Hot Start Every Cycle
M T TS ATG TMg Pfu

14. Polymerase Thermostability
DNA Polymerase Polymerization Opt Half Life 95oC
Pfu oC 19 hours
Taq oC 1 hour

15. Characteristics of PfuTurbo
Improved version of Pfu
Same high fidelity as Pfu
Higher PCR product yield than Pfu alone
Can use 1 min/kb extension times
Works well with low amounts of DNA template

16. PCR Enhancing Factor (PEF)
Isolated from Pyrococcus
Lacks DNA polymerase and exonuclease activities
Information on identity/mechanism of action is proprietary

17. PfuTurbo vs Other Polymerases
M PT P V DV Pw K T M PT P V DV Pw K T M
5.2 kb
0.9 kb

18. PfuTurbo vs Pfu
M T P T P T P T P T P
8 kb
6.1 kb
5.2 kb
1.9 kb
0.9 kb

19. Characteristics of TaqPlus Precision
Blend of Taq and Pfu
High Yield
High Fidelity
Length
PCR Buffer
optimized for both yield and fidelity

20. Accuracy of Polymerase Blends in PCR

21. PCR with TaqPlus Precision vs Taq
Taq TaqPlus Precision
M M kb

22. Characteristics of TaqPlus Long
Blend of Taq and Pfu
Long targets
Fidelity similar to Taq
Two optimal buffers supplied
high salt buffer helps prevent smearing in long target amplification
low salt buffer

23. PCR with Taq vs TaqPlus Long

24. Qualities of Taq Taq2000 is the recombinant Taq
Native Taq is also available
Taq is trusted by customers
Taq has been well studied

25. TaqExtender The Pfu component of TaqPlus Long
Can be added to any Taq to make a polymerase blend

26. Perfect Match PCR Enhancer
Taq native Taq M

27. Opti-Prime PCR Optimization Kit
12 buffers
matrix of pH, Mg, KCl
Adjuncts
DMSO, formamide, BSA, glycerol, ammonium sulfate, Perfect Match
Recommended for use with Taq

28. Related PCR products RoboCycler Nucleotides and primers
gradient
Nucleotides and primers
StrataPrep PCR purification kit
Cloning Vectors
PCR-Script Kits
Affinity protein expression kit
QuikChange site-directed mutagenesis kit
Cyclist DNA sequencing kits