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IVT mRNA/circRNA Synthesis, SPRI Purification & qPCR Quantification

URL: https://cloud.ginkgo.bio/protocols/ivt-rna-synthesis-qpcr Status: Ginkgo Certified Price: $99/sample Turnaround: up to 12 business days (extended: up to 17 business days for longer or clonal DNA constructs) Throughput: 384-well format throughout

Overview

End-to-end automated RNA production and quantification pipeline ("data in / data out"). Submit DNA sequences via CSV; Ginkgo sources DNA synthesis from commercial providers (e.g., Twist), then runs PCR template prep, in vitro transcription (linear mRNA or circRNA), SPRI bead purification, and qPCR quantification.

Note: The circRNA pipeline is limited to Permuted Intron-Exon (PIE) with group I introns.

Input

Output

Automated Workflow

Phase 1 - PCR Template Preparation

  1. Transfer primers (Labcyte Echo)
  2. Transfer templates (Labcyte Echo)
  3. Stamp mastermix (Bravo 384)
  4. PCR thermocycle (ATC384)

Phase 2 - In Vitro Transcription

  1. Stamp mastermix (Bravo 384)
  2. IVT incubation (ATC384)
  3. Stamp DNase mix (Bravo 384)
  4. Circularization for circRNA (ATC384)

Phase 3 - SPRI RNA Purification

  1. Load decks with reagents (Bravo 384)
  2. Magnetic bead purification (Bravo 384)
  3. Unload decks (Bravo 384)

Phase 4 - Quantify RNAs

  1. Stamp RNAs (Bravo 384)
  2. RNA yield / purity quantification (Lunatic)

Phase 5 - Circularization Quantification

  1. Echo qPCR setup (Labcyte Echo)
  2. Stamp mastermix (Bravo 384)
  3. Run RT-qPCR (CFX Opus)

Ordering

Use Cases