Working Copy: What is the difference between LUTHOR HD 3' mRNA-Seq and QuantSeq 3’ mRNA-Seq?

LUTHOR utilizes a pre-amplification step.
LUTHOR libraries are therefore generated from amplified RNA molecules.
The pre-amplification step 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.

QuantSeq utilizes a different chemistry where no pre-amplification step is employed, requiring 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.

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

The pre-amplification initial IVT amplifies the original mRNA molecules and step enables the generation of libraries from ultra-low input RNA, down to the single-cell level.

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

RNA amplification in LUTHOR HD is templated by the endogenous mRNA molecules instead of using cDNA intermediates.

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

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.