Photographic recording of fluorescent DNA bands on agarose gels.

cloning could be achieved by using this quick, alternative protocol. Not only does this Na3VO4-inhibition procedure save time, but also note that in this protocol the adjustment of vector-to-insert ratio in the ligation reaction is not necessary. To ligate with the CIAP/Na3VO4-treated vector (ca. 50 ng), we recommend using as much excess insert as is practical. Eliminating the quantitation step for both vector and insert further simplifies the process and reduces the hands-on time of the cloning procedure. In summary, the procedure we describe in this report should make the subcloning method using alkaline phosphatase not only a less costly but also a time-effective procedure. A high percentage of recombinants could be quickly achieved without using any special cloning vector or bacterial strain. We feel that this protocol is especially useful for cloning procedures having no appropriate screening method (e.g., blue/white colony selection) or no background reduction strategy [e.g., digesting the ligation product before transformation with a selection enzyme that cuts the vector between the two cloning sites (3,4)].