| TA
Cloning |
Perform
TA cloning with SimVector |
The TA cloning
method takes advantage of the terminal
transferase activity of some DNA polymerases
such as Taq polymerase. This
enzyme adds a single, 3'-A overhang
to each end of the PCR product. This
makes it possible to clone this PCR
product directly into a linearized
cloning vector with single, 3'-T overhangs.
To clone such PCR product the linearized
cloning vector with single, 3'-T overhangs
called a T- vector are used. The PCR
products with dA overhang, are mixed
with this vector in high proportion.
The complementary overhangs of "T"
vector and PCR product will get hybridized
and ultimate joining of two DNA fragments
accomplished by T4 DNA ligase. |
| |
| Perform
TA cloning with SimVector |
SimVector
automatically generates graphical
DNA sequence maps of designed recombinant
DNA molecules displaying vector -
donor relationships and the corresponding
cloning protocol. Use commercial T-vector
or design your own T-vector using
restriction digestion and recombine
with the donor in sense or anti-sense
orientation to generate highly accurate
recombinant DNA sequence map.
|
| TA
Cloning: Design linearized commercial
T Vector |
|
| TA
Cloning: Design donor
vector |
|
| TA
Cloning: Recombinant vector |
|
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