PCR
using Dynazyme Taq
WHEN DO YOU
USE DYNAZYME?
When you want to PCR a fragment and you need the amplicon to be perfect.
For example when building a construct, difficult or long PCR templates,
or preparing something for sequencing.
Don’t use Dynazyme for any ‘normal’ PCR, just because
your PCR isn’t working (try temperature gradient or magnesium
gradient for that), for example SNIP SNP mapping or to test the presence
of a mutation. Only when you need the product for sequencing, or for
an injectable construct.
Dynazyme is a lot more expensive that Promega Taq, so don’t try
it for the first time in 40 –50 ?l reactions, 10 or 20 ?l reactions
work just as well.
General
MIX
| Buffer (Mg added) |
2 µl |
|
| Dynazyme |
1 µl |
(units per µl
might change with lot) |
| dNTPs |
2 µl |
|
| Primer F |
0.5 µl |
|
| Primer R |
0.5 µl |
|
| DNA |
1 µl |
(optimize) |
| H2O |
to 20 µl |
|
The buffer is also supplied
without Mg, with a separate vial of Mg, if you need to adjust the concentration.
|
|
|