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  • suppression subtractive hybridization and 454

    Has anyone done suppression subtractive hybridization PCR (SSH) on cDNA prior to 454 sequencing? Any tips, protocols, etc would be greatly appreciated.

  • #2
    Originally posted by AKroxy View Post
    Has anyone done suppression subtractive hybridization PCR (SSH) on cDNA prior to 454 sequencing? Any tips, protocols, etc would be greatly appreciated.
    Probably not what you are looking for, but the following uses an Evrogen kit to normalize cDNA libraries prior to 454 sequencing:

    http://www.pubmedcentral.nih.gov/art...?artid=2689275

    Evrogen also sells SSH reagents, so that could be a place to look.

    Phillip

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    • #3
      ssh + 454

      Hello,
      Do you know if are there some information about supression substrative hubridization using clontech kit prior to pyrosequencing 454???
      Thanks in advance!!!

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      • #4
        I tried tweaking that protocol to get rid of homopolymers so we could use the samples in 454 sequencing, but I failed. I would have needed to spend a lot more time on it to get it to work.

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