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  • Complete ligation adaptor sequences

    Hello everyone,

    I am producing Ion Torrent sequencing libraries both using the ligation based library kit and the ampliseq strategy. The only adaptor sequences I can find from Lifetech are the ones involved in ampliseq:

    Forward primer (Primer A-key):
    5’-CCATCTCATCCCTGCGTGTCTCCGACTCAG-template-specific-sequence-3’
    Reverse primer (Primer P1-key):
    5’-CCTCTCTATGGGCAGTCGGTGAT-template-specific-sequence-3’

    For quantitation purposes I use the Taqman based qPCR kit. This is however only possible with the ligated libraries. On this basis I assume the double stranded oligos used for ligation contains a longer sequence, than the one generated in the ampliseq PCR.

    Does anyone have this sequence of the ligation adaptors?

    I know this may well, for some obscure reason, be one of Lifetechs little secrets and that I may have to go through the trouble of cloning and sequencing the adaptors myself.

  • #2
    Exactly what I was looking for! Thank you very much genseq!

    Comment


    • #3
      Hi Apex,

      The sequences of the fragment library adaptors can be found in Appendix D (pp. 45) of the current Ion Xpress™ Plus gDNA Fragment Library Preparation User Guide, as available on the Ion Community.

      Ion AmpliSeq™ Custom Panels may be designed using the Ion AmpliSeq™ Designer website.


      Originally posted by Apex View Post
      Hello everyone,

      I am producing Ion Torrent sequencing libraries both using the ligation based library kit and the ampliseq strategy. The only adaptor sequences I can find from Lifetech are the ones involved in ampliseq:

      Forward primer (Primer A-key):
      5’-CCATCTCATCCCTGCGTGTCTCCGACTCAG-template-specific-sequence-3’
      Reverse primer (Primer P1-key):
      5’-CCTCTCTATGGGCAGTCGGTGAT-template-specific-sequence-3’

      For quantitation purposes I use the Taqman based qPCR kit. This is however only possible with the ligated libraries. On this basis I assume the double stranded oligos used for ligation contains a longer sequence, than the one generated in the ampliseq PCR.

      Does anyone have this sequence of the ligation adaptors?

      I know this may well, for some obscure reason, be one of Lifetechs little secrets and that I may have to go through the trouble of cloning and sequencing the adaptors myself.

      Comment


      • #4
        adaptor sequences for Ion S5

        Hi,
        I'm new to Life Tech semi conductor Sequencing and only have MiSeq experience. We were offered a demo run of the Ion S5 and I would like to produce the library beforehand with our custom protocol.

        Can anyone tell me if the Ion torrent adaptors are the same for the Ion S5?

        Fwd: CCATCTCATCCCTGCGTGTCTCCGACTCAG

        Rev: CCTCTCTATGGGCAGTCGGTGAT

        cheers,
        JoF

        Comment

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