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      Bio-Rad SEQuoia Stranded RNA Library Prep Kit

      Q: How do I adapt the Bio-Rad SEQuoia Complete or Express Stranded RNA Library Prep Kit to sequence on the G4 Sequencing Platform?

      A: Follow the Bio-Rad SEQuoia Express Stranded RNA Library Prep Kit or SEQuoia Complete Stranded RNA Library Prep Kit protocol, but set up PCR amplification with Singular Genomics (SG) UDI primers in the Library Amplification (Indexing PCR) step instead of the primers listed in the protocol. After library prep, deplete the library from fragments derived from ribosomal RNA with the SEQuoia RiboDepletion Kit.

      LIBRARY PREPARATION PROTOCOL

      Perform the following steps:

      1. Perform one of the following Bio-Rad SEQuoia protocols, but use the following PCR reagents for the Library Amplification (Indexing PCR) step:
      Reagent
      Volume
      SG UDI Primers (5 µM each)
      2.5 µL
      SEQuoia Express Amplification Mix (green cap)
      or
      Reagent F Amplification Mix (green cap)
      25 µL
      cDNA library sample
      22.5 µL
      Total Volume
      50 µL
      1. Perform the depletion described in the SEQuoia RiboDepletion Kit protocol
      2. Make sure to perform the library Quality Control steps listed here: Amplified Library Prep Quality Control

      DEMONSTRATED G4 SEQUENCING EXPERIMENTS

      For demonstrated SEQuoia Stranded RNA sequencing experiments on the G4, see the App Note RNA Sequencing on the Singular Genomics G4 Sequencing Platform Using SEQuoia Library Preparation Chemistries .

      ORDERING INFORMATION

      SG Indexed PCR primers:

      Part #
      Product
      SG UDI Primers (1-96)
      SG UDI Primers Set A (1-24)
      SG UDI Primers Set B (25-48)

      Bio-Rad SEQuoia Stranded RNA Library Prep Kit:

      Part #
      Product
      Bio-Rad SEQuoia Express Stranded RNA Library Prep Kit, 24 rxn
      Bio-Rad SEQuoia Express Stranded RNA Library Prep Kit, 96 rxn
      Bio-Rad SEQuoia Complete Stranded RNA Library Prep Kit, 24 rxn
      Bio-Rad SEQuoia Complete Stranded RNA Library Prep Kit, 96 rxn

      Bio-Rad SEQuoia RiboDepletion Kit:

      Part #
      Product
      Bio-Rad SEQuoia RiboDepletion Kit, 24 reactions

      Version: 27 Oct 2023