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  • Anyone know what the fill-in enzyme is?

    Does anyone know the enzyme Roche uses for the fill-in reaction in the gsflx library prep kit? Anyone know an enzyme that does the same thing?

    This is to fill-in a phosphodiester bond on one strand after adaptors are added.

    Thanks!

  • #2
    have a look at the methods section in this paper
    i've used this protocol successfully. step 14 is the adapter fill-in step using Bst polymerase.

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    • #3
      Thanks, Marklar. I will check it out.

      Comment


      • #4
        Bst polymerase is used in that paper.

        Can I use Large Fragment of DNA Polymerase I (Klenow Fragment) instead? It is said to have strand displacement property.

        Thanks for any comments.

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        • #5
          Yes, I think you can use the 3'- 5' exo- Klenow Fragment. Check out NEB's latest flyer. I am told the Bst polymerase adds dAs.

          Good luck!

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          • #6
            Thanks gaster!

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