sequencing chromosomal DNA

Beth Rogers erogers at umbc.edu
Wed Dec 7 12:18:15 EST 2005


We are trying to sequence chromosomal DNA using ABI's BigDye sequencing 
protocols and are getting no signals (plasmid and PCR sequencing works 
fine).  We have upped the DNA and primer amounts still with no luck. 
We are thinking of trying longer denaturation times(i.e. 10 mins), 
shearing, NaOh denaturation, DNAse nicking...  Anyone have any 
experience with this that could offer some suggestions?  Thanks, ejr


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