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  • sajoshi
    Member
    • Dec 2015
    • 26

    How to get library size

    Do my libraries look OK? Are they overloaded. How to I get the library size, I cant seem to adjust the upper and lower marker. Attached is the bioanalyzer profile. I called Agilent and they said clean the pins. Everytime I call them its the same answer.

    Please help me
    Thanks,

    Note: TruSeq Total RNA RiboZero kit was used
    Attached Files
  • Ancestro
    Member
    • Oct 2015
    • 15

    #2
    It seems that you are carrying over some impurities in the library. Some of them (the last ones) maybe are sub-fragmented inserts also. Maybe you should make another bead purification and retry to run a chip.

    If Agilent tells you to clean the pins, just to be curious, do you made that also? Sometimes happens the same and we clean them with pure water poured in the transparent chip around 30 min before using it, if it is the case, you can prepare another blue matrix, it might be old and is sensible to light.

    In the bioanalyzer there is an option to convert time (s) to pb, according to the molecular marker peak. The button is in the view menu.

    Your average insert size might be between 200 and 300 pb



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    • nucacidhunter
      Jafar Jabbari
      • Jan 2013
      • 1250

      #3
      JL-9.3 WT and FG-9.6 MUT are OK. Other libraries have been over amplified. All samples except JL-9.3 WT have been overloaded.

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