Background in In Situ Hybridizations

[Reese Bolinger]

Hi:

We have been trying to work up a protocol for performing in situ 
hybridizations in Xenopus embryos following the protocol of Harland 
(1991) and have been having problems with both probe and antibody 
associated background.  Specifically, we are wondering:

1.  Do you need to use lamb serum (which we have been) or is sheep serum 
ok?

2.  To reduce the probe-related background, should you RNAse longer or 
with a higher RNAse concentration?

Any other suggestions you have on reducing background would also be 
greatly appreciated.

                                                  Thanks!

                                                  Reese