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Reagent Preparation Guidance

Reagent Preparation

Monarch Carrier RNA

For reconstitution of Carrier RNA based on kit size used, add 125 µl (NEB #T4010S) or 750 µl (NEB #T4010L/X) nuclease-free water, invert or pipette to mix, and transfer to an RNase-free microfuge tube. Keep on ice. Prepare single-use aliquots and store at –20°C. Avoid multiple freeze-thaw cycles.

Lysis Buffer Bead Mix

  • Prepare Lysis Buffer Bead Mix immediately before use (i.e., immediately before Sample Lysis, or during the Sample Lysis, Proteinase K incubation step).
  • Prepare Lysis Buffer Bead Mix by combining Monarch StabiLyse DNA/RNA Buffer (included), Monarch Carrier RNA (included), Isopropanol (user supplied), and Monarch Mag Beads M1 (included) as described in the protocol, adding components in the order listed. Vortex magnetic beads ~20 seconds immediately before use to form a homogeneous suspension. Carefully open the bottle after vortexing to ensure the magnetic beads do not spill.
  • Store Lysis Buffer Bead Mix at room temperature.
  • If preparing a master mix, prepare excess to ensure sufficient volume of the mixture for each reaction. We recommend an overage of up to 15%.

User-Prepared Wash Buffers

Viral DNA/RNA Wash Buffer: Prepare Viral DNA/RNA Wash Buffer in a user-supplied tube or bottle (free of nucleases) by combining Monarch Buffer BX (included), nuclease-free water (included), and Isopropanol (user supplied) as described in the protocol, adding components in the order listed. The nuclease-free water included in the kit is intended to be used for preparing this wash buffer. Users are instructed to prepare an amount of wash buffer that includes up to 15% excess to ensure sufficient volume for each reaction. Prepare Viral DNA/RNA Wash Buffer fresh, as a precipitate may form upon storage. 

80% Ethanol: Prepare 80% fresh ethanol using 100% ethanol (user supplied) and nuclease-free water (user supplied) in a user-supplied bottle, free of nucleases. Users are instructed to prepare an amount of 80% ethanol that includes up to 15% excess to ensure sufficient volume for each reaction. To support our sustainability efforts, we have not included additional bottles and nuclease-free water for these 80% ethanol wash steps to avoid shipping excessive materials that may not apply to all users.