HELP! F.I.S.H. FAILING!

pallderdice at kean.ucs.mun.ca pallderdice at kean.ucs.mun.ca
Thu Jun 1 11:41:07 EST 1995


 I HAVE RECENTLY BEGAN THE USE OF F.I.S.H. FOR MAPPING PROBES 
 SIZES 1 TO 50 kb( PLASMIDS, COSMIDS, YACS ). I AM USING 
 BIO & DIG LABELING(BLR & BM ) KITS, G-50 COLUMNS, AND ONCOR REAGENTS 
 AND PROTOCOLS. THE RESULTS FOR THE PAST FEW YEARS(BY OLD RESEARCHER)
 WHERE 10/700 SLIDES THAT WORKED. I AM NOW SEARCHING FOR THE OPTIUMN PROTOCOL 
 TO GIVE SUCESSFUL AND CONSISTENT RESULTS. THE AREAS MOST CONCERNED 
 WITH ARE; SLIDE PRETREATMENT, POST HYB WASH, AVIDIN & ANTIBODY WASHES.    
 ANY OTHER HELPFUL HINTS WOULD BE WELCOMED.

 ALSO: DO YOU USE PROTEINASE K AND OR RNAse H OR A?
       DID YOU EVER TRY 0.1M TRIETHANOLAMINE AND 0.25% 
       ACETIC ANHYDRIDE TREATMENT OF SLIDES?

THANK YOU,
GLEN EDMUNDS




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