Insertion of unique sites into YAC arms for rapid physical analysis following YAC transfer into mammalian cells

Method for rapid restriction analysis of YAC clones.

The development of the yeast artificial chromosome (YACs) technology (3) has provided, by virtue of its capacity to carry large fragments of genomic DNA, the possibility of linking the genetic map and the physical map of an organism. This requires the alignment of overlapping YAC clones into contigs for which a number of strategies have been described as follows: the use of sequence tagged site...

A method for high efficiency YAC lipofection into murine embryonic stem cells.

We describe a modified protocol for introducing yeast artificial chromosomes (YACs) into murine embryonic stem (ES) cells by lipofection. With a decreased DNA:cell ratio, increased concentration of condensing agents and altered culture conditions, this protocol reduces the requirement for YAC DNA to a few micrograms, improves the recovery of neomycin-resistant ES colonies and increases the yiel...

Enhancement of Pyrimidine K562 and Yac-1 Cells Nucleoside Uptake into by Cadeguomycin

Cadeguomycin markedly stimulated the uptake of thymidine, deoxycytidine and uridine into the acid-insoluble fraction of K562 human leukemic cells, but did not significantly affect adenosine incorporation. The enhancement of pyrimidine nucleoside uptake was 617 fold over the control. Aspartate incorporation into nucleic acid was not significantly blocked by the antibiotic, suggesting that the st...

Preparation of YAC end fragments from the whitehead/MIT mouse YAC library pRML vectors.

Physical interactions between DNA and sepiolite nanofibers, and potential application for DNA transfer into mammalian cells

Nanofibers of sepiolite, a natural silicate belonging to the clay minerals family, might constitute a potential promising nanocarrier for the non-viral transfer of bio-molecules. We show here that sepiolite nanofibers efficiently bind different types of DNA molecules through electrostatic interactions, hydrogen bonding, cation bridges, and van der Waals forces. Moreover, Fourier-transform infra...