EzPCR™ BASIC 5x 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 5x 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 5x PCR Premix is recommended for use in conventional PCR process.

  As Premix is supplied as a 5x concentration format, users should adjust a final reaction to 1x (if final reaction volume is 20 ㎕, 4 ㎕of 5x Master Mix should be added).

  One unit of rTaq DNA polymerase is contained in 4 ㎕ 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 5x Premix	4 ㎕
Primer (10 pmol/㎕)	each 0.5 ㎕
Template DNA	0.1 - 50 ng
PCR grade distilled water	-㎕
Add distilled water to make 20 ㎕ final volume.

 

 

2. Set PCR cycling as follows

 

Initial denature at 95 ℃ : 3 min
		< 1 kbp	> 1 kbp
Denature	95 ℃	10 - 30 sec	30 sec
Anneal	Tm ℃	10 - 20 sec	30 sec
Extend	72 ℃	10 - 30 sec	1 min / Kbp

 

   * Set 25-35 PCR cycles for effective amplification.