Taq DNA Polymerase Comparison (Part 2)
TransBionovo vs Qiagen vs NEB vs Biobasic Taq DNA PolymeraseTaq DNA Polymerase Comparison…which one should you choose?
Nowadays, many suppliers offer Taq DNA Polymerases. Our aim was to perform a Taq DNA Polymerase Comparison. Our Taq DNA Polymerase Comparison assay consisted in PCR amplification of a fragment of the human β-globin gene from purified human genomic DNA or mouse genotyping of DsRED, Cre or flox/src from unpurified mouse gDNA. PCR and DNA electrophoresis were performed with the miniPCR and blueGel systems respectively.
Details
Date: June 16th, 2016
Type of experiment: PCR from human gDNA
DNA Polymerase: EasyTaq DNA Polymerase
Competitors: NEB, Sigma, Qiagen, BioBasic
PCR reaction setup for β-globin amplification:
- H2O : fill to 20 ul
- 10x buffer : 2 ul
- dNTPs (2.5 mM): 1.6 ul (0.2 mM final)
- Forward primer (10 uM): 0.4 ul (200 nM final)
- Reverse primer (10 uM) : 0.4 ul (200 nM final)
- gDNA 10 ng/ul : 2 ul
- Taq DNA Polymerase (5 u/ul) : 0.2 ul
- (BioBasic Taq required the addition of MgSO4 - as recommended)
PCR cycling for β-globin amplification :
- Denaturation: 60s at 94 °C
- 35 x
- Denaturation: 15s at 94 °C
- Annealing: 15s at 59 °C
- Extension: 35s at 68 °C
- Final extension: 300s at 68 °C
Taq DNA Polymerase Comparison : NEB M0267 vs Qiagen Taq DNA Polymerase
Amplification of different fragment sizes of the human β-globin gene using BEN Taq M0267 or Negaiq Taq DNA Polymerase.
Similarly to EasyTaq in above figure, both BEN and Negaiq Taq DNA Polymerases performed well and were specific in this Taq DNA Polymerase Performance Comparison.
NOTE: Sigma’s RedTaq and NEB’s OneTaq both failed in the majority of the PCR reactions for b-globin (data not shown)
Which Taq should you choose for mouse genotyping by PCR?
This assay consists in PCR amplification of DsRED, Cre or flox/src from unpurified mouse gDNA..
Details
Client: None
Date: June 16th, 2016
Type of experiment: PCR from human gDNA
DNA Polymerase: EasyTaq DNA Polymerase, TransTaq HiFi DNA Polymerase (buffer I)
Competitors: NEB, Sigma, Qiagen
PCR reaction setup for DsRed amplification:
- H2O : to 20 ul
- 10x buffer : 2 ul
- dNTPs (2.5mM): 1.6 ul (0.2 mM final)
- F1 (10 uM): 0.4 ul (200 nM final)
- F2 : 0.4 ul
- R1 : 0.4 ul
- R2 : 0.4 ul
- gDNA : 1 ul unpurified gDNA
- Polymerase (5 u/ul) : 0.2 ul
PCR cycling for DsRED amplification :
- Denaturation: 60s at 94 °C
- 35 x
- Denaturation: 15s at 94 °C
- Annealing: 20s at 57 °C
- Extension: 10s at 72 °C
- Final extension: 300s at 72 °C
PCR reaction setup for Cre and flox/src amplification:
- H2O : to 20 ul
- 10x buffer : 2 ul
- dNTPs (2.5mM): 1.6 ul (0.2 mM final)
- F1 (10 uM): 0.4 ul (200 nM final)
- R1 : 0.4 ul
- gDNA : 1 ul unpurified gDNA
- Polymerase (5 u/ul) : 0.2 ul
PCR cycling for DsRED amplification :
- Denaturation: 60s at 94 °C
- 35 x
- Denaturation: 15s at 94 °C
- Annealing: 20s at 63 °C
- Extension: 15s at 72 °C
- Final extension: 120s at 72 °C
DsRed Genotyping - Taq DNA Polymerase Comparison
Amplification of the DsRED transgene using EasyTaq, TransTaq HiFi, BEN Taq or Negaiq Taq DNA Polymerase.
Conclusion
After evaluating the performance of Transgen Biotech (now TransBionovo) EasyTaq® DNA Polymerase in PCR amplification of various targets from human or mouse genomic DNA, performed on purified or crude DNA extracts, we conclude that in most PCR assays and using that same amount of Taq units, EasyTaq was equivalent to competitor Taq DNA Polymerases. However, EasyTaq was clearly superior at performing DNA amplification of difficult targets.
Approximative cost units/dollar (CAD):
EasyTaq® DNA Polymerase AP111-02 : 3000u / 255$ = 11.76 u/$
Qiagen Taq 201207 : 5000u / 2357$ = 2.12 u/$
NEB Taq M0267X : 4000u / 620$ = 6.45 u/$
BioBasic 9K-001-0033: 6000u / 750$ = 8 u/$
Best Performance + Best Price = EASY PCR!
Want us to Compare your DNA Polymerase before you make the Switch?
Put us in charge of optimizing PCR conditions for you. It’s FREE !
DNA Polymerase Comparison Chart
Specifications and pricing of commercially available Taq DNA Polymerases and High-Fidelity DNA Polymerases in Canada.
Detection Limit of High-Fidelity DNA Polymerases on gDNA
Published on Jan 30, 2017 by Simon Roy, Ph.D Detection Limit of High-Fidelity DNA Polymerases using Human gDNA for Template We previously determined the detection limit of different Taq DNA Polymerases using human genomic DNA as a template for PCR. For this instance,...
PCR Purification Kit Comparison - DNA Clean Up
Which is the Best Kit for DNA Clean Up? Our aim was to perform a PCR Purification Kit Comparison from different vendors. At Civic Bioscience, we distribute the Favorprep™ and EasyPure® product lines from FAVORGEN and TransBionovo respectively. First, the FavorPrep™...
miniPCR Review : How I use a portable PCR thermocycler to help life scientists day-to-day.
The miniPCR and blueGel systems represent the core instruments involved in the assistance we provide to real scientists who conduct life science research in Canada. Both laboratory instruments were essential in solving dozens of problems that prospect customers were...
GC-rich PCR: Comparison of FastPfu FLY and Q5 High-Fidelity DNA Polymerases
GC-rich PCR: FastPfu FLY vs Q5® High-Fidelity DNA Polymerase GC-rich PCR can be difficult to achieve Our aim was to verify compare FastPfu FLY (TransBionovo) and Q5 (NEB) high-fidelity DNA Polymerase at amplifying a GC-rich sequence (human ARX; 79% GC) from purified...
Detection Limit of Taq DNA Polymerase
What's the Detection Limit of Taq ? How many DNA copies from gDNA? There are MANY Taq DNA Polymerases out there on the market. Some are driven by strong marketing and others by strong performance. Our aim was to verify the Detection Limit of Taq DNA Polymerases in PCR...
PCR Cycling: High-Fidelity DNA Polymerase Comparison
PCR Cycling comparison between TransBionovo’s TransStart® FastPfu and FastPfu FLY with competitor High-Fidelity DNA Polymerases. Finding a reliable and versatile high-fidelity enzyme remains a challenge in DNA research. Our goal was to extend the data presented in our...
High-Fidelity DNA Polymerase Comparison using unpurified gDNA
High-Fidelity DNA Polymerase Comparison on difficult genomic DNA samples. Modern commercially available high-fidelity DNA Polymerases are derived from either Pfu and KOD. Pfu is an enzyme found in the hyperthermophilic archaeon Pyrococcus furiosus. KOD is an enzyme...
Trackbacks/Pingbacks