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  • vtorda
    Junior Member
    • Oct 2016
    • 4
    #1

    SmartSeq cDNA validation with TapeStation

    Hi All,

    I have RNA extractions from fungal fruiting body tissue, produced by cryo-sectioning-->LCM-->PicoPure RNA extraction kit.
    I have started to use SmartSeq V4 to produce cDNA. After the amplification of the cDNA we checked the cDNA profile by TapeStation.

    But the profile doesn’t look like as SmartSeq manual suggests: we have a peak around 400bp instead of 2500bp.
    Anybody has experienced cDNA profile different from the profile in SmartSeq manual?
    The company who will do sequencing said that this profile is similar what they usually see on cDNA produced by SmartSeqV4.
    What is your opinion?

    Thank you all!!
    Attached Files
    Tags: cdna synthesis, single cell analysis, smartseq2
    • nucacidhunter
      Jafar Jabbari
      • Jan 2013
      • 1250
      #2
      It looks fine. I would clean it with 0.6x beads before shearing and library prep for RNA-Seq.
      Last edited by nucacidhunter; 11-23-2016, 12:52 PM.

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