In situ RNA-RNA probes

[yungyungbee]

hi, i am new to use in situ hybridization


i made in situ RNA probes from PCR template and T7 RNA polymerase, the produced probes has 1.5ug/ul concentration. i loaded 3 ul of the sample to a normal TBE DNA gel, and the gel photo show a smear with 2 sharp bands....one band is the size of my template and the other band is larger....is the smear due to too much probes loaded??....or sth wrong in the process of producing the probe??

i would like to know whether i have succesffuly produce the probe or not....and should i dilute the probe before doing the in situ hybridization? and can it be simply diluted DEPC-treated water??.....


thanks a lot