EzPCR™ 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. EzPCR™ 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’ to 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 ℃.
EzPCR™ 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
EzPCRTMBASIC 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.