genome size?

[Glen Shearer]

Michael Cooley (szcooley at chip.ucdavis.edu) wrote:
: What is the easiest method of determining genome size. I really only need 
: an estimate, I don't want to do CoT analysis is I can avoid it. Seems 
: like I remember some way which involves Southern blots.


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One quick way:  genomic reconstruction.
  1.  Apply two-fold dilutions of genomic DNA to membrane w/ slot blot.
  2.  Do same with single copy gene from your organism
  3.  Denature, bake, & probe blot with 32-P labeled same gene.
  4.  Quantitate the band densities w/ laser scanner or image analysis
      from the autorad.
  5.  Since you know the exact amount of DNA in each slot you
      can calculate how much genomic DNA you need to give the
      same intensity as x fg single copy gene and thus calculate
      amount of genomic DNA to give one copy of the gene hence
      haploid genome size.

 Regards,
 Glen
 [gshearer at whale.st.usm.edu]