Reagents

PacBio reagent kits are available for different workflow steps emphasizing the flexibility of the system. From small PCR amplicons to DNA libraries of 10's of kilobases, multiple DNA samples can be sequenced in an individual SMRT Cell or the same sample can be sequenced across multiple SMRT Cells according to your experiment design and data needs. This customizable run capability provides for cost efficiencies in both large and small sequencing projects.

The PacBio System has two main reagent components: the front-end template preparation and the on-instrument DNA sequencing. All SMRTbell template preparations undergo similar procedures.

Front-End Template Preparation
The SMRTbell template preparation and DNA polymerase binding steps may be performed manually at the benchtop or by an automated liquid-handling system.

  1. Target Enrichment and Multiplexing

  2. SMRTbell Template Preparation Reagent Kits
    • DNA and cDNA samples are converted into a primer-annealed SMRTbell library format.


  3. DNA Polymerase Binding Reagent Kits
    • Primer-annealed SMRTbell templates are bound to a DNA polymerase in preparation for sequencing.

On-Instrument DNA Sequencing
The DNA sequencing step is performed on the PacBio System. Sequencing reagent kits supply all materials needed (other than plastic disposables) for easy instrument loading.

  1. DNA Sequencing Reagent Kits
    • Supports automated, on-instrument DNA sequencing with all required chemistry and reagents. Used in conjunction with SMRT Cell 8Pacs.


Sample prep kits

The Barcoded Universal F/R Primer Plate – 96 contains both forward and reverse primers to be used in a PCR reaction to add barcodes to PCR amplicons which have been amplified with the universal sequence tags. Each well contains sufficient reagents for a single use. To be used in conjunction with the SMRTbell Template Prep kit.

The SMRTbell Template Prep Kit 1.0 contains reagent supplies to perform SMRTbell library preparations of non size-selected and size-selected primer-annealed SMRTbell libraries for insert sizes ranging from 500 bp to over 20 kb. Reagent quantities support 10 library preparations.

The SMRTbell Barcoded Adapter Complete Prep Kit – 96 contains both the SMRTbell Barcoded Adapter Prep Kit and Barcoded Adapter Plate - 96. Reagent quantities support 96 individual Barcoded Adapter End-Repair and Ligation reactions. To be used with SMRTbell Damage Repair Kit to completely prepare primer-annealed SMRTbell libraries.

The SMRTbell Damage Repair Kit contains reagents to perform DNA damage repair and anneal the sequencing primer. Reagent quantities support 10 preparations.

The SMRTbell HT Template Prep Kit contains reagent supplies to perform 96 automated SMRTbell library preparations of non size-selected, primer-annealed SMRTbell libraries for insert sizes ranging from 500 bp to over 20 kb. Reagents come in 4 x 24 reaction sets. The reagents in the SMRTbell HT Template Prep Kit are found in different volume portions to the SMRTbell Template Prep Kit 1.0 and are not scalable from one to another.

The reagents are compatible with protocols that have been codeveloped for the Agilent NGS Workstation Option B and the Sciclone NGSx Workstation from PerkinElmer. The available protocols for these systems have been validated with the PacBio 10 kb Template preparation workflow.
To learn more, please contact your local Field Application Scientist (FAS) or Sales Representative.

AMPure PB magnetic beads are specifically formulated for the SMRT sequencing workflow. Beads are used for DNA cleanup and purification during sample extraction and at various stages in the workflow.



Binding kits

The DNA/Polymerase Binding Kit P6 v2 contains reagent supplies to bind prepared DNA template libraries to the DNA polymerase P6 in preparation for sequencing on the PacBio RS. Reagent quantities support 24 binding reactions. DNA Polymerase P6 should be used only in conjunction with Sequencing Reagent 4.0.

The MagBead Kit contains reagent supplies to capture DNA polymerase and template complex for loading onto the PacBio RS. Kit components include: MagBeads and MagBead Buffer kit.



Sequencing reagents and kits

The DNA Sequencing Kit 4.0 contains a sequencing reagent plate with C4 chemistry and OS Enzyme for Single-Molecule Real-Time sequencing on the PacBio RS. Reagent quantities support 8 sequencing reactions to be used in conjunction with SMRT Cell 8Pac(s) and DNA Polymerase P6.

The DNA Sequencing Bundle 4.0 contains reagent and disposable supplies for 80 reactions of Single-Molecule Real-Time sequencing. It contains: 10 DNA Sequencing Kits 4.0, 1 SMRT Cell Oil pack, 1 Sequencing Plate Septa pack, 1 Tube Septa pack.



Sequencing Controls

The DNA Internal Control Complex contains a fixed template of 2 kb length, bound to DNA Polymerase P6 for use as an internal sequencing control. Reagent quantities provide spike-in controls for a minimum of 24 samples.