Template Preparation Kits

The TPK 2.0 Bundle contains one kit of each of the following: SMRTbell Express Template Prep Kit (TPK) 2.0 (PN: 100-938-900), SMRTbell Enzyme Clean Up Kit (PN: 101-746-400) and Sequencing Primer v2 (PN: 101-847-900). The TPK 2.0 bundle builds off of the SMRTbell Express TPK 2.0 workflow -- a single-tube, addition-only strategy to minimize handling-induced DNA damage for quick library generation. The SMRTbell Enzyme Clean Up kit helps maximize intact SMRTbell templates. Library generation can be completed as few as 5 hours, excluding size selection. SMRTbell templates can be used to generate large-insert HiFi reads for variant detection or de novo assembly.

Each kit contains sufficient reagents to prepare 9 HiFi libraries using 10 ug of gDNA per preparation.

Note: Sequel and Sequel II System compatible. Kit components are sold separately.

• Resource: Procedure & Checklist - Preparing HiFi SMRTbell Libraries Using SMRTbell Express Template Prep Kit 2.0
• Resource: Procedure & Checklist - Using AMPure PB Bead for Size Selection
• Resource: Procedure & Checklist – Preparing 10 kb Library Using SMRTbell Express Template Prep Kit 2.0 for Metagenomic Shotgun Sequencing
• Product insert: SMRTbell Express Template Prep Kit 2.0
• Product insert: SMRTbell Enzyme Clean Up Kit
• Product insert: Sequencing Primer v2
• SDS: SMRTbell Express Template Prep Kit 2.0
• SDS: SMRTbell Enzyme Clean Up Kit
• SDS: Sequencing Primer v2

The SMRTbell Express Template Prep Kit 2.0 (TPK 2.0) supports insert sizes from 250 bp to >50 kb. This kit employs a single-tube, addition-only strategy to minimize handling-induced DNA damage for library generation in as little as 4 hours, excluding size selection. We recommend this kit for most applications except for shotgun metagenomics (use TPK 1.0, PN: 100-259-100) or large-insert HiFi reads are required. To generate large-insert HiFi reads, the SMRTbell Enzyme Clean Up Kit (PN: 101-746-400) and Sequencing Primer v2 (PN: 101-847-900) are also needed which can be purchased together in the TPK 2.0 Bundle (PN: 101-843-100).

Each kit contains sufficient reagents to prepare either 18 standard reaction large-insert libraries, 18 Iso-Seq libraries, 48 multiplexed microbial genomes, or 96 reactions for amplicon sequencing.

Note: Sequel and Sequel II Systems compatible.

• Resource: Product Note: Large-insert genomic libraries with SMRTbell Express Template Prep Kit 2.0
• Resource: Product Note: Multiplexed Microbial Genome Sequencing with SMRTbell Express Template Prep Kit 2.0
• Resource: Procedure & Checklist - Preparing gDNA Libraries Using the SMRTbell Express Template Preparation Kit 2.0
• Resource: Procedure & Checklist – Preparing SMRTbell Libraries Using Express Template Prep Kit 2.0 With Low DNA Input
• Product insert: SMRTbell Express Template Prep Kit 2.0
• SDS: SMRTbell Express Template Prep Kit 2.0

The SMRTbell gDNA Sample Amplification Kit is designed to amplify sample starting from ~5-20 ng of gDNA to a sufficient quantity for template preparation procedures using the SMRTbell Express Template Prep Kit 2.0.

Each kit contains sufficient reagents to amplify 18 samples.

Note: Compatible with the Sequel II System

• Resource: Procedure & Checklist - Preparing HiFi SMRTbell Libraries from Ultra-Low DNA Input
• Resource: Application Note Considerations for Using the Low and Ultra-low DNA Input Workflows for Whole Genome Sequencing
• Product insert: SMRTbell gDNA Sample Amplification Kit
• SDS: SMRTbell gDNA Sample Amplification Kit

The Iso-Seq Express Oligo Kit contains two oligonucleotides to support the Iso Seq Express workflow used in conjunction with the SMRTbell Express Template Prep Kit 2.0 and NEBNext Single Cell/Low Input RNA Library Prep Kit. The oligonucleotides are the PacBio Template Switching Oligo (TSO) and PacBio cDNA PCR primer.

Reagents support 24 RNA transcriptome library preparation.

Note: Sequel and Sequel II Systems compatible.

• Resource: Procedure & Checklist: Iso-Seq Express Template Preparation for Sequel and Sequel II Systems
• Product insert: Iso-Seq Express Oligo Kit
• SDS: Iso-Seq Express Oligo Kit

No-Amp Accessory Kit contains Adapter Annealing Buffer, Sequencing Primer v4, and 10X Primer Buffer v2. Annealing buffer is to be used during SMRTbell Adapter hairpin annealing. Sequencing Primer v4 and 10X Primer Buffer v2 is used to anneal Sequencing Primer v4 to final SMRTbell templates.

Reagent quantities support up to 20 SMRTbell Express TPK 2.0 library preparations using standard input amounts.

Note: Sequel and Sequel II Systems compatible. Pacific Biosciences does not sell a kit for carrying out the overall No-Amp Targeted Sequencing method. Use of this method may require rights to third-party owned intellectual property.

• Resource: Procedure & Checklist – No-Amp Targeted Sequencing Utilizing the CRISPR Cas9 System
• Product insert: No-Amp Accessory Kit
• SDS: No-Amp Accessory Kit

AMPure PB beads are used for 1) DNA cleanup and purification during various stages in the SMRTbell library template preparation workflow, 2) buffer exchanges, 3) template concentration, 4) clean up of excess polymerase and primer from polymerase-bound SMRTbell complexes, and and 5) size selection.

Reagent quantity supplied at 5 mL volume.

Note: Compatible with all PacBio Systems

• Resource: Procedure & Checklist – AMPure PB Bead Purification of Polymerase Bound SMRTbell Complexes
• Resource: Procedure & Checklist - Using AMPure PB Bead for Size Selection
• Product insert: AMPure PB
• SDS: AMPure PB

The Elution Buffer (50 mL) is used with the SMRTbell library template preparation and barcoding associated kits for workflows requiring additional elution buffer and AMPure clean-up steps.

Reagent quantity supplied at 50 mL volume.

Note: Compatible with all PacBio Systems

• Product insert: Elution Buffer (50 mL)
• SDS: Elution Buffer (50 mL)