Biomedical Engineering Reference
In-Depth Information
8. The POE-PCR product was a kind of sticky solution and can
be used to transform competent cells directly. Do not try to
clean the POE-PCR product with a DNA cleaning kit. The
DNA multimers may be stored at 4 °C for several days for
future transformation. Do not freeze the POE-PCR product
since it caused severe precipitation. It was frequent to see some
high molecular weight DNA at the position of more than
10 kb instead of DNA multimers, which were intermediates
generated by POE-PCR.
9. Do not add too much POE-PCR product in competent cells
for transformation, since the high-salt PCR solution could
decrease the transformation effi ciency [
15
].
Acknowledgments
This work was supported mainly by the Shell GameChanger Program,
DOE BioEnergy Science Center, DOE ARPA-E Petro, and the
Virginia Tech CALS Biodesign, and NSF SBIR I grant to PZ.
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