PCR cloning in pDK101 XcmI, reference

Russell L. Malmberg russell at dogwood.botany.uga.edu
Thu Aug 17 07:07:51 EST 1995

"Direct Cloning of DNA Fragments Generated by PCR"
Kovalic, D., Weisblum, B.
1995, Methods in Neurosciences, volume 26:255-261
Sarkar, G., ed., Academic Press

I learned of this review article containing suggested protocols for use
of the vector pDK101 with the enzyme XcmI to clone PCR fragments,
and thought I would pass the reference along.

Some notes from the article:
- pDK101 is available from ATCC in strain number 77406
- pDK101 has XcmI sites that leave 3' unpaired Ts on the vector,
     following removal of a 16 base pair stuffer fragment.
- they recommend gel purification of the vector away from the 16 bp fragment
- they recommend the addition of a kinase step just before adding ligase

Thanks to the authors for pointing this reference out to me.

Russell L. Malmberg
russell at dogwood.botany.uga.edu

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