EzPCRTM BASIC 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,
bromophenol blue, and stabilizer). PCR can be performed simply by adding primer
pair and template.
EzPCRTM
BASIC 5x PCR Master Mix 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. EzPCRTM BASIC 5x PCR Master Mix is
recommended for use in conventional PCR process.
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).
One unit of rTaq DNA polymerase is contained
in 4 ㎕ of 5x Master Mix.
QC tests
Performance test, exo/endo
nuclease contamination test, stability test. genomic DNA contamination test.
Storage Condition
Store
at -20 ℃.
EzPCRTM BASIC 5x PCR Master Mix is stable
for at least 1years at recommended storage condition
Standard Protocol
1. Prepare the 20 ㎕ PCR solution as
follows
EzPCRTM BASIC 5x Master Mix | 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.
