Q] Digestion,Elution,Ligation

Hyunmin Kim vurk at hymail.hanyang.ac.kr
Fri Jun 19 02:38:22 EST 1998

I'm doing

   1.Enzyme digestion, 2 Elution of  digested DNA fragment , 3.
Dephosphorylation,and 4. Ligation

      I did the whole step in          1->2->3->4..order.
         However what if I do   in  1->3->2->4.

   What is more efficient ?
      #1.Dephosphorylatiion before Elution  or #2. Dephosphorylation
after Elution


    I did Elution by using BioRad Elution KIT. and I found that about
1/10(or less than 1/10) DNA was eluted.
    The efficiency is too low. Is this low efficiency is in general?
    In addition, I  heard that the eluted DNA is hard to be ligated by
DNA ligase.
    Do you have any idea about these?

Hyunmin Kim
Laboratory of Microbiology
Department of Biology
College of Natural Science
Hanyang Univ.
Haengdang-dong 17                Fax   : 082-02-290-0952
Seoul, South Korea               E-mail: vurk at hymail.hanyang.ac.kr

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