uv crosslinking of dna to filters

suter at VAX.MPIZ-KOELN.MPG.DBP.DE suter at VAX.MPIZ-KOELN.MPG.DBP.DE
Thu Dec 19 07:19:04 EST 1991


dear netteers,

we generally crosslink our filters as follows:
260 nanometers of uv / 75 seconds / intensity: 2000 micro Watt per
square centimeter of filter.

this was optimized for genescreen, however works for nitrocellulose also.

filters of both kinds were hybridized up to 8 times, some loss of signal
did occur (30%?).

we have tested this for several time intervals: 70 or 80 of uv irradiation
already showed a "dramatic" loss of signal strength: say 10%.

((with 70 or 80 i mean seconds naturally))

however ! we have now switched to zetaprobe from biorad, which functions
better in our hands. as far as i know it can not be used for plaque lifts.

ciao, clemens



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