EzPCR™ BASIC 2x PCR Premix is a ready-to-use premix containing all the components essential for a PCR and agarose gel electrophoresis (DNA polymerase, dNTP, reaction buffer, glycerol, bromophenol blue, and stabilizer). PCR can be performed simply by adding primer pair and template.
EzPCR™ BASIC 2x PCR Premix contains rTaq DNA polymerase that is a recombinant thermostable DNA polymerase from Thermus aquaticus. rTaq DNA polymerase has a 5’ à 3’ exonuclease activity and generates a 3’dA overhang. EzPCR™ BASIC 2x PCR Premix is recommended for use in conventional PCR process.
As Premix is supplied as a 2x concentration format, users should adjust a final reaction to 1x (if final reaction volume is 20 ㎕, 10 ㎕ of 2x Master Mix should be added).
One unit of rTaq DNA polymerase is contained in 10 ㎕ of 5x Premix.
Application
- Appropriate for standard PCR amplifications.
- Designed to perform PCR amplification on all DNA templates.
- Amplification of cDNA and genomic DNA
- Colony PCR
QC tests
Performance test, exo/endo nuclease contamination test, stability test. genomic DNA contamination test.
Stroage condition
Store at -20 ℃.
EzPCR™ BASIC 5x PCR Premix is stable for at least 1years at recommended storage condition.
Features
- 5’ → 3’ exonuclease : yes
- 3’ → 5’ exonuclease : no
- strand displacement : no
Standard Protocol
1. Prepare the 20 ㎕ PCR solution as follows
EzPCR™BASIC 2x Premix
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4 ㎕
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Primer (10 pmol/㎕)
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each 0.5 ㎕
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Template DNA
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0.1 - 50 ng
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PCR grade distilled water
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-㎕
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Add distilled water to make 20 ㎕ final volume.
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2. Set PCR cycling as follows
Initial denature at 95 ℃ : 3 min
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< 1 kbp
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> 1 kbp
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Denature
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95 ℃
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10 - 30 sec
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30 sec
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Anneal
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Tm ℃
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10 - 20 sec
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30 sec
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Extend
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72 ℃
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10 - 30 sec
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1 min / Kbp
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* Set 25-35 PCR cycles for effective amplification.