yeast chromosome isolation

Anthony Tomlinson tomlinson at pplros.demon.co.uk
Tue Mar 14 05:10:56 EST 1995


In article <ab83f7390002100401f1@[128.231.218.92]> Michael Lichten,
lichten at HELIX.NIH.GOV writes:
>>I am preparing to isolate yeast chromosomes for the purpose of CHEF
>>analysis and was wondering about good protocols to use for doing so.  I
>>have the basic protocol from Maniatis.  Does this work well or are there
>>some "secret" tricks that need to be done to ensure success.  Also, the
>>protocol suggests using zymolyase 5000.  Can one substitute zymolyase
>>100,000 for this or would that be detrimental?  Any tidbits of info are
>>welcome.
>>
>You can dispense with zymolyase completely and get very nice chromosomes.
>A protocol was published which basically says that you can skip straight to
>the sarkosyl/proteinase K step but I cannot find the reference.  Anyone out
>there?
>

I think you're referring to this one:

				 Ibeas and Jimenez (1993). Nuc Acid Res  21:3902.



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