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  • Ion Ampliseq library prep query

    Hi everybody,

    This is my first post here. I am really puzzled with respect to some of my library profiles I am currently seeing in my Ion Ampliseq library preparation with the Ion Ampliseq Transcriptome Human Gene expression kit. (Cat# A26326). We use very low quality RNA (degraded as well as low conc., can't help it as our samples are like that). But we get nice good peaks even if using these RNAs. We usually use the SpeedVac to concentrate our RNAs before doing the prep.

    This is how the library looks like in my hands:

    Click image for larger version

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    But recently, I have been getting really weird profile of the libraries. Please see below:

    ​​​​​​Click image for larger version

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    Both of these are from Std RNA. As seen in second figure, there are 3 peaks which do not look like libraries and the conc. is also very low. Above 1000pg/ul for first profile and less than 200 pg/ul for second profile. I am not undersatnding what those 3 peaks are and why this happened.
    Please help.

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