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Taq DNA Polymerase Comparison (Part 2)

TransBionovo vs Qiagen vs NEB vs Biobasic Taq DNA Polymerase

Taq 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 :

  1. Denaturation: 60s at 94 °C
  2. 35 x
    • Denaturation: 15s at 94 °C
    • Annealing: 15s at 59 °C
    • Extension: 35s at 68 °C
  3. Final extension: 300s at 68 °C

Taq DNA Polymerase Comparison : EasyTaq vs BioBasic Taq DNA Polymerase

Amplification of different fragment sizes of the human β-globin gene using EasyTaq or BioBasic (Feldan) Taq DNA Polymerase.

EasyTaq performed much better than BB Taq in our Taq DNA Polymerase Performance Comparison assay.

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 :

  1. Denaturation: 60s at 94 °C
  2. 35 x
    1. Denaturation: 15s at 94 °C
    2. Annealing: 20s at 57 °C
    3. Extension: 10s at 72 °C
  3. 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 :

  1. Denaturation: 60s at 94 °C
  2. 35 x
    1. Denaturation: 15s at 94 °C
    2. Annealing: 20s at 63 °C
    3. Extension: 15s at 72 °C
  3. 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.

Cre and flox/src Genotyping - Taq DNA Polymerase Comparison

Amplification of Cre or flox/src from transgenic mice gDNA samples using EasyTaq, BEN Taq M0267 or Negaiq Taq DNA Polymerase.

EasyTaq performed better than BEN and Negaiq Taq DNA Polymerases.

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!

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