in situ hybridization

Heather Etchevers etchever at lovelace.infobiogen.fr
Mon Dec 9 07:25:59 EST 1996


Are you sure there is not a simple typo?

We put 100 microliters of probe diluted in hyb buffer on an ordinary microscope
slide. Given if you are isolating the section with a PAP or wax pen you
can get away with less, in fact 60 microliters is really sufficient. But
10 seems very little. After making the probe and resuspending it isn DTT
then I use 10 microliters + 90 of buffer. Then I coverslip each slide
with a 22x50mm slip and not bothering to use nailpolish, put it in a chamber
humidified with 50% formamide, 2xSSC on paper towels in the bottom. I
haven't had any problems with drying out. Also,m don't try to pry up the
coversplips afterward, but rather let them soak in your first rinse bath
15 minutes, then replace this bath and continue. The slips sually will 
fall off by themselves and your sections will suffer less.

Heather




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