DNA Prep for
microinjection
The following 2
kits are the most popular methods for preparing DNA for pronuclear
microinjection:
QIAquick Gel Extraction Kit (70 bp – 10 kb)
QIAEX II Gel Extraction Kit (40 bp – 50 kb)
With both kits,
use T-lowE (10 mM TRIS, 0.1 mM EDTA)
which is available from the Transgenic Core for the final elution step. You may make your own T-lowE however
the stock available from the Core has been embryo tested and is recommended.
Even with kits, we see
variation in the purity of the DNA submitted. The purity of the DNA and the accuracy of the quantification
are the 2 most important factors in the success of your injections. Even when using a kit, careful
attention to details is essential!
For those who are opposed to
the use of kits, here is an additional protocol: