Q] Digestion,Elution,Ligation

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


Hi~
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.

Question1.>>
   What is more efficient ?
      #1.Dephosphorylatiion before Elution  or #2. Dephosphorylation
after Elution

Question2.>>

    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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