High Density DNA array protocol needed!

Csaba Kiss csakis at ki.se
Thu Mar 22 11:09:56 EST 2001


Hi!
I would like to prepare high density grids of soem 1000  PCR products. They
are derived from a subtracted cDNA library. The clones of the cDNA libraries
were  PCRed up with two common flanking primers. We have the BIOMEK 2000
robot in the lab with a high density replica tool. I used this tool before
to array bacteria on nylon filters, and grew them overnight on LB plates. I
noticed that the sizes of the obtained colonies were very variable, and the
filter-to-filter colony variation was also quite high. That's why i decided
that next time I will array DNA directly on the nylon filters. I am looking
for protocols or tips&hints how to make the best of it. Of course I want to
have equal amounts of DNA spots on each filter and between the filters the
corresponding spots should also have the same amount of DNA.
I am looking for any advice. For example is it better to do the arraying
with wet nylon filter or dry? Is DNA arraying better or worse than bacterium
arraying.

Best regards:
                        Csaba Kiss





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