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In Situ hybridization suddenly stopped working!!

RNA in situ hybridization ish

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#1 Tara Burns

Tara Burns

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Posted 24 September 2015 - 04:52 AM

Hello everyone, 

 

Well I'm pretty stumped, I just started doing in situ which is not a common technique in our lab.  The first couple of times everything went great, the third time the staining was specific but just lighter.  This time there was absolutely no staining.  I haven't changed the technique, tissue, probe, etc.  Nearly everything I have is aliquoted in the freezer and I used all new aliquots this week.  The only solutions that aren't aliquoted are the ethanols, PBS/DEPC, DEPC water, and the ingredients for the NTMT.  Is this most likely RNAse contamination??  What would you do to troubleshoot.  I guess I will start with dumping all my solutions that aren't aliquoted?  SOOOOO FRUSTERATED!!







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