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Simple DNA-Seq library preparation kit enables library construction in three steps. NuQuant library quantification technology included providing accurate library molarity in seconds.
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Tab 01 / Overview
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In addition to our standard Celero products, we offer customizable Celero solutions for high throughput DNA-seq, high throughput amplicon sequencing, and 384 multiplexing with NuQuant technology.
Please speak with an NGS Expert to find out more.
The Celero DNA-Seq library preparation kit is an innovative system designed to help researchers streamline library preparation for Illumina sequencers. Celero features a fast, easy-to-use, addition-only workflow that eliminates post-ligation bead purification. Also included is Tecan's breakthrough library quantitation method, NuQuant®, that saves both time and cost in measuring library concentration for pooling. With the proven DimerFree® technology you can use from 10 ng to 500ng of intact DNA with a single kit without the worry of GC bias.
NuQuant app for Qubit® can be downloaded here on Github.
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Tab 02 / Highlights
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Celero DNA-Seq has the simplest and shortest workflow. Master mixes are provided at each step and bead purifications are eliminated after ligation step for an easy, streamlined library preparation.
Celero provides superior sequencing efficiency of high and low GC organisms. gDNA from E. coli (51 % GC), R. sphaeroides (69 % GC) and S. aureus (33 % GC) were mixed to equal mass and sequenced with Celero DNA-Seq and kits from Companies K and I. The percentage of each genome is given as a fraction of total reads. PCR amplified Celero libraries demonstrate the same bacterial balance as the PCR-free control. Company K has a significant bias against the A/T-rich genome
Celero DNA-Seq provides reduced base bias. 10 ng of an equal mass mixture of Covaris sheared gDNA from E. coli (51 % GC), R. sphaeroides (69 % GC) and S. aureus (33 % GC) was sequenced with Celero DNA-Seq and kits from Companies K and I. Graphs from FastQC showing the base distribution (first 25 bases shown in graph; A - Green, T - Red, C - Blue, G - Black) demonstrate that Company K has a significant footprint in the first 10 bases and a high A/T bias. Company I also has a noticeable footprint in the first 8 bases.
NuQuant has the lowest variability for library quantitation. A set of 8 libraries was distributed to 6 different users at multiple institutions. Users were asked to use NuQuant and their own preferred method of library quantitation (number denoted by n). For each library, and each quantification method, the percent coefficient of variation (%CV) of molar concentration was calculated. NuQuant produced the lowest variation, followed by qPCR. Bioanalyzer gave the highest variation.
Strong correlation between NuQuant molarity and read numbers. Two users prepared 8 libraries each from 10 ng of genomic DNA sheared using a Covaris and amplified either 10 or 13 cycles to produce both lower and higher library concentrations. All measurements were performed in duplicate, across two users, for a total of 16 libraries. Purified libraries were quantified by NuQuant, then equal volumes of all libraries were pooled and sequenced on one lane of an Illumina sequencer.
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Tab 03 / Specifications
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Specification |
Specification/Description |
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Compatible Platform |
Compatible Platform Illumina HiSeq, MiSeq, NextSeq, NovaSeq |
Starting Material |
Starting Material Purified DNA, cDNA |
Input Amounts |
Input Amounts 10 ng - 500 ng |
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Tab 04 / Faq
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Celero DNA-Seq includes all necessary buffers, primers and enzymes for library construction. SPRI purification beads and EvaGreen are not included.
Required equipment includes a microcentrifuge, pipettes, vortexer, a thermal cycler, a magnetic plate for 0.2 mL tubes, strips, or plates and a fluorometer. An Agilent Bioanalyzer or Fragment Analyzer may also be useful for optional analytical tests. A comprehensive list of required and recommended equipment can be found in Section II.B of the User Guide.
Celero DNA-Seq is an end-to-end solution designed to generate libraries for Illumina sequencing starting from gDNA or cDNA and has not been tested with alternative library preparation systems.
We recommend a column-based extraction method, including: Qiagen QIAprep Miniprep or DNeasy Miniprep kits, Zymo Quick-DNA kits and Thermo Fisher PureLink Genomic DNA kits.
We do not recommend the use of these methods as any carryover of organics may inhibit downstream enzyme activity. If using, we recommend using a column-based purification of the downstream enzyme activity. If using, we recommend using a column-based purification of the DNA prior to input into the kit.
Celero DNA-Seq has been designed for use with a broad range of different organisms. Special consideration should be given when using low-input samples from organisms with large genomes.
We strongly recommend using high quality DNA with the A260:A280 ratio in excess of 1.8. Use of DNA samples with lower ratios may result in low library yield. This kit is designed for use with DNA samples of high molecular weight with little or no evidence of degradation. FFPE DNA may be compatible with the PCR workflow. When using degraded samples, we recommend using inputs of 200 ng or greater. While it is impossible to guarantee satisfactory results with all degraded samples, this system may work with many samples that are moderately degraded.
Yes.
No.
RNAClean XP beads are certified to be RNase and DNase free. We have tested both RNAClean XP and AMPure XP beads in our kits and observe no difference in performance between products.
Due to the large number of commercially available magnets, we do not have a comprehensive list of compatible products. However, many magnets are compatible. As long as the magnet is strong enough to clear the solution of magnetic beads, it can be applied to the system. We have the following guidelines for selecting a magnetic separation device:
Please refer to section V. H. for guidelines on quantitative and qualitative assessment.
The adaptors add 136 bp to the library.
Celero DNA-Seq libraries are compatible with Illumina sequencing platforms.
Please follow manufacturer’s recommendations for library QC, quantification, balancing and loading of the amplified library on the sequencer.
Celero DNA-Seq libraries are designed for use with the standard Illumina sequencing primers for both single-end and paired-end sequencing applications.
Yes. The libraries produced using this kit can be used for both single-end and paired-end sequencing. Special consideration should be given to the expected insert size in the paired end assay. The workflow generates libraries with an average insert size that is dependent on the fragmentation conditions used.
Each barcode is a minimum edit distance of 3 from any other barcode. This means that a minimum of three edits (replacement, insertion, or deletion) must occur before one barcode becomes a different barcode. For further details on the barcode design strategy, please refer to Faircloth BC, Glenn TC (2012), Not All Sequence Tags Are Created Equal: Designing and Validating Sequence Identification Tags Robust to Indels. PLoS ONE 7(8): e42543. doi:10.1371/journal.pone.0042543.
The final Celero libraries can be analyzed using standard pipelines. To remove adaptor artifacts use standard TruSeq sequences for read trimming.
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Tab 05 / Literature
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Robust DNA-Seq library prep kit for a broad range of sample types starting with as little as...
DNA-Seq library prep kit with optimization free enzymatic fragmentation for a 3 step...
Simple and robust enzymatic PCR-free solution for DNA-Seq library preparation in less than 2.5 hours.
For research use only. Not for use in diagnostic procedures.