Description

 

  EzPCRTM XO 5x PCR Master Mix is a ready-to-use premix containing all the components essential for a PCR and agarose gel electrophoresis (DNA polymerase, dNTP, reaction buffer, glycerol, phenol red and stabilizer). PCR can be performed simply by adding primer pair and template.

 

  HiPi Plus DNA polymerase in Master Mix is suitable for a high fidelity amplification of DNA fragments. This enzyme is designed for a reliable amplification of complex targets with a robust yield and high specificity. HiPi plus DNA polymerase is recommended for use in conventional PCR that products are less than 5 kb (for lambda DNA).  HiPi Plus DNA polymerase generates a mixture of PCR products with blunt end and 3’ -dA overhangs.

 

  As Master Mix 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). 

Applications

 

 - Long PCR

 

 - RT-PCR

 

 - Amplification of cDNA and genomic DNA

 

 - High fidelity PCR for a cloning 

 

Qualifying Test

 

  Performance test, exo/endo nuclease contamination test, stability test. genomic DNA contamination test. 

 

Storage Condition

 

  Store at -20 ℃.

 

  EzPCRTMXO 5x PCR Master Mix is stable for at least 1years at recommended storage condition. 

 

Features

 

 - 5’ → 3’ exonuclease : no

 

 - 3’ → 5’ exonuclease : weak

 

 - strand displacement : no 

 

Troubleshooting

 

 

 

1. No products

 

 - Confirmyourtemplateisintact:Try another reaction

 

  with a result assured primer pair and templates.

 

 

 

2. Smear bands or smeared background

 

 - Reduce template concentration: High concentration of template can lead to smearing of PCR products. Generally, 1 pg - 10 ng of plasmid DNA  and 10-100 ng of genomic DNA are working well

 

 - Increase annealing temperature

 

 - Set up a reaction mix on ice

 

 

 

3. Non-specific bands

 

 - Increase annealing temperature

 

 - Consider using PCR additives, like 1-2% DMSO or 0.5 - 1 x Q Buffer

 

 - Confirm specificity of your primers

 

 - Adjust concentration of template

 

 

 

4. Low yield

 

 - Increase PCR cycle number

 

 - Be sure appropriate concentration of your template is added.