Posted 30 November 2011 - 05:05 AM
Thanks to all!!!
Posted 30 November 2011 - 08:47 AM
If you are concerned about cross hybridization, do a blast search -- if you find long hits (50+bp) of very high homology (very few mismatches), you could clone segments of your gene that exclude those regions.
Hyb temperature is related to your hyb buffer. The protocol above is done at 50deg, which is common when substantial amounts of formamide are included in the buffer.
Edited by David C H, 30 November 2011 - 08:51 AM.