SMARTer Apollo library prep system overview
Unrivaled results with simplified, automated NGS library preparation
Next-generation sequencing (NGS) applications like DNA-seq, RNA-seq, and ChIP-seq enable researchers to delve deeper into the human genome. Although NGS applications allow diverse and detailed analyses, NGS library preparation is encumbered by extensive hands-on and labor-intensive workflows. To address this, the SMARTer Apollo system provides a complete, walkaway solution with validated protocols and optimized chemistries to produce consistent libraries for DNA-seq, RNA-seq, and ChIP-seq applications
- Convenient library prep—reduce hands-on time with walkaway automation
- Flexible throughput—run any batch size up to 96 samples per run
- Optimized inputs—conserve precious samples
- Reliable results—increase reproducibility and minimize contamination with a completely enclosed, automated system
Flexible, streamlined workflow
The SMARTer Apollo system offers best-in-class performance and flexibility. Manual NGS library prep workflows are labor-intensive, occupying researchers for long periods of hands-on time. The SMARTer Apollo system frees researchers from manual steps by automating the critical portions of the workflow, reducing hands-on time to just 30 minutes, depending on the sample throughput and script used. Automated NGS library prep on the SMARTer Apollo system allows researchers to perform other experimental or analysis tasks during NGS library preparation.
Flexibility on the SMARTer Apollo system enables researchers to easily adapt to variable sample throughput needs without wasting reagents. In addition to both DNA- and RNA-seq library prep protocols, this versatile system efficiently performs related protocols for sample preparation and cleanup, such as poly A+ RNA enrichment, rRNA depletion, and PCR cleanup.
Optimized input amounts
Sample input amount and quality are important considerations for any sequencing application. Committing too much of your sample to a workflow or getting a low yield means wasted material, delayed experiments, poor results, or insufficient sequencing material. SMARTer PrepX kits and related scripts resolve these issues by generating comparable, high-quality NGS libraries regardless of input concentrations within a defined range (Tables I and II).
Input amount | 10 ng | 100 ng |
---|---|---|
Adapter reads (%) | 0 | 0 |
Confidently mapped reads (%) | 96.61 | 96.43 |
Repetitive reads (%) | 1.84 | 1.80 |
Low-quality reads (%) | 0 | 0 |
Table I. Reliable sequencing coverage and performance. High-quality libraries were prepared using the SMARTer PrepX DNA library prep kit on the SMARTer Apollo system from either 10 ng or 100 ng of E. coli genomic DNA. Libraries were sequenced on the Illumina MiSeq® platform (2 x 150 bp).
SMARTer PrepX kits provide every reagent necessary for library prep in prealiquoted, color-coded, single-use reagent strips. Each SMARTer PrepX chemistry is compatible with a script with variable throughputs and minimum input requirements, as summarized below.
Cat. # | SMARTer Apollo reagent kit | Compatible script | Script throughput | Minimum input amount per sample |
---|---|---|---|---|
640101 | SMARTer PrepX Complete ILMN DNA Library Kit, 24 Samples | PrepX ILM 8 | 1–8 | 1 ng DNA |
640102 | SMARTer PrepX Complete ILMN 32i DNA Library Kit, 96 Samples | PrepX ILM 32i | 4–32 | 1 ng DNA |
640096 | SMARTer PrepX RNA-Seq for Illumina Library Kit, 24 Samples | PrepX mRNA 8 | 1–8 | 100 pg polyA-enriched RNA; 2 ng rRNA-depleted mRNA |
640097 | SMARTer PrepX RNA-Seq for Illumina Library Kit, 48 Samples | PrepX mRNA 48 | 6–48 | 100 pg polyA-enriched RNA; 2 ng rRNA-depleted mRNA |
640098 | SMARTer PrepX PolyA mRNA Isolation Kit, 96 Samples | PrepX PolyA 8 or PrepX PolyA 48 | 1–8 or6–48 | 100 ng total RNA |
640099 | SMARTer PrepX Small RNA for Illumina Library Kit, 24 Samples | PrepX small RNA 8 | 1–8 | 100 ng total RNA |
640100 | SMARTer PrepX Small RNA for Illumina Library Kit, 48 Samples | PrepX small RNA 48 | 6–48 | 100 ng total RNA |
Table II. Input amounts required for various combinations of SMARTer PrepX kits and preprogrammed SMARTer Apollo scripts.
Maximize yield and consistency
Manual NGS library preparation is time-consuming and subject to human error, affecting the quality of your data. Consistency is key when processing multiple samples, and the SMARTer Apollo system, combined with SMARTer PrepX chemistries and scripts, allows you to create consistent libraries without variation caused by manual pipetting. Comparison of manual and automated poly A+ RNA enrichment on the SMARTer Apollo system demonstrates that automation delivers higher poly A+ RNA yield and diversity as compared to the manual method (Figure 1).
Figure 1. Poly A+ RNA enrichment on the SMARTer Apollo system produces better quality and yield as compared to manual poly A+ RNA enrichment. Panel A. Manual poly A+ enrichment was performed on 1 µg of human brain total RNA in triplicate. Bioanalyzer traces of these manual replicates show variation in yield. Panel B. Automated poly A+ RNA enrichment was performed on 1 µg of human brain total RNA and a no-template control using the SMARTer Apollo system with the SMARTer PrepX PolyA mRNA Isolation Kit, 96 Samples. Automated enrichment (Panel B, red trace) results in higher poly A+ RNA yield and diversity as compared to manual enrichment (Panel A).
SMARTer PrepX chemistries have been validated to be a seamless solution for consistent sample preparation, cleanup, and NGS library prep (Figure 2). Many of these protocols employ in-tip bead separation for efficient capture of libraries in a highly reproducible manner. Learn more about in-tip bead separation on our application page.
Figure 2. Consistent library size with a SMARTer PrepX DNA library kit. Eight technical replicate E. coli libraries were created with a SMARTer PrepX DNA library kit on the SMARTer Apollo system, demonstrating consistent post-PCR library sizes.
Minimize waste
Most SMARTer PrepX reagents are aliquoted into disposable strips for single use in a variety of sample configurations, eliminating pipetting error and excess waste. To make our validated protocols faster, easier, and error-free, the reagents are color-coded and accommodate a variety of sample throughputs.