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LUTHOR HD

  • LUTHOR utilizes a pre-amplification step.
    LUTHOR libraries are therefore generated from amplified RNA molecules.
    The pre-amplification step HD utilizes an initial in vitro transcription (IVT) step before the production of cDNA and its amplification.

  • The initial IVT amplifies the original mRNA molecules and enables the generation of libraries from ultra-low input RNA, down to the single-cell level.RNA amplification in LUTHOR is templated by the endogenous mRNA molecules instead of using cDNA intermediates

  • LUTHOR HD includes a 12nt UMI sequence located at the beginning of Read 2, introduced during oligo(dT) priming.

  • LUTHOR HD requires asymmetrical paired end sequencing to read out the UMI.

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QuantSeq FWD

  • QuantSeq utilizes a different chemistry where no pre-amplification step is employed, requiring and therefore requires higher RNA input amounts.

  • QuantSeq captures the 3' end of transcripts by oligo(dT) priming and reverse transcription.
    During reverse transcription, a partial Illumina compatible adapter is introduced at the 3' end. After removal of the template RNA, the second strand is generated by random priming and extension, using a DNA polymerase.

 
  • QuantSeq is best sequenced with single read sequencing.