Genomic DNA/RFLP

tom siepmann tsiepman at post.its.mcw.edu
Sun Nov 12 14:57:12 EST 1995


Greetings all,

My advisor wants me to prove that a cell line of ours that is acting
strangely (strange in a good way) is actually a subcloned cell of the original
parent line (newly) acquired from ATCC.  He wants me to prove this 
via RFLP on the genomic DNA from each (clonal) cell line.

I have a method that involves suspending the cells in agarose and then
digesting away all but the DNA...Has anyone else had success with this
method or could recommend a method of his/her own?  I haven't been
able to get any DNA...I don't know if this is due to using a poor
method or my inexperience/ineptitude in working with DNA.  People I
ask for advice tell me to send the cells out to have this done.  Am
I trying to do the impossible?  If the DNA gets sheared then that 
screws up the RFLP...the marker probe will hybridize to fragments of
different size therefore making it seem like the cells are truly 
of different makeup when they may not actually be.

Any help/guidance is appreciated.

Sincerely,

Jennifer Potter
humble grad student
Medical College of WI (Go Packers!)
jras at post.its.mcw.edu



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