How can I extend my sequencing results?

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    • #14387
      sahart
      Participant

      Hi all,

      I have sequenced around 400 bp of a cDNA, I am wondering how can i extend my sequencing result to whole the cDNA, kindly noted that I have no information about other part of this mRNA.

      Thank you, 🙂

      Sahar

    • #103188
      magicsiew
      Participant

      carried out RACE-PCR

    • #103197
      canalon
      Participant

      Sequence walking: Design a new primer at the end of the one you have.

    • #103204
      sahart
      Participant

      Thanks all

    • #103205
      sahart
      Participant
      quote canalon:

      Sequence walking: Design a new primer at the end of the one you have.

      How much should I add cDNA as template?

    • #103206
      canalon
      Participant

      Exact same reaction, new sequencing primer, you move a few hundred bp forward until you reach the end. Long but that is what used to be done in the olden days before shotgun sequencing. And if you have access to a good sequencing facility, where read length of above 700bp are the norm that makes things even faster.

    • #103210
      JackBean
      Participant

      if we talk about one read, there is not much what you can do as a customer. Just provide enough high-quality DNA and primers which anneal fine.

    • #103229
      magicsiew
      Participant
      quote canalon:

      Sequence walking: Design a new primer at the end of the one you have.

      Hi, if I need to elongate the 5′ direction?

    • #103234
      JackBean
      Participant

      just design reverse complement primer 😉

      But I think, that chromosome walking is basically the same as RACE-PCR 😉

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