ted-CapALL™ Library Preparation Kit

$350.00$3,360.00

The ted-CapALL™ kit converts DNA and RNA molecules simultaneously into next generation sequencing (NGS) libraries. The robustness of the ted-CapALL™ workflow allows the analysis of picograms of nucleic acids directly from limited samples (5pg – 100ng) or cell lysates (> 10 cells).  The concurrent analysis of DNA and RNA allows cell type identification by linking gene expression data to chromatin accessibility profiles. The ted-CapALL™ Kit generates sequencing libraries for Illumina® sequencing platforms.

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Description

For Illumina sequencing systems

Cat. No. Format Storage Kit stability
TR-1012-08 8 reactions per kit -20°C 1 year from the date of manufacture
TR-1012-96 96 reactions per kit -20°C 1 year from the date of manufacture

The ted-CapALL™ Kits generate sequencing libraries for Illumina® sequencing platforms. The novel “ted” technology used in these kits generate gene expression data as well as nucleosomal coverage data directly from cell and tissue homogenates providing orthogonal data to correlate gene expression with nucleosomal epigenetic profiles. ted-CapALL™ protocols do not require ribosomal RNA removal as rRNA is converted into library molecules at substantially low efficiency.

Attributes:

Input sample 1 - 10000 cells lysate
5 pg - 100 ng total RNA or mRNA
Total nucleic acid in medium or serum
Input amount 7 uL of extracted RNA
6 uL of lysates with 1 to 10000 cells
2 uL of cell medium or serum
Data produced Sequencing reads delineating nucleosomal-free regions of the genome.
Sequencing reads for coding and non-coding RNA with < 10% of total reads mapping to ribosomal RNA.
Data processing Q20 filter, trim adaptors, remove <20bp reads and duplicates
seqtk trimfq -q 0.01 $Read2 ; with bioinformatic tool seqtk from https://github.com/lh3/seqtk.
Protocol time
  1. Lysate or homogenate to library: 80 minutes (1 tube reaction)
  2. Indexing PCR & QC → 1 hr
No bead cleanup until after indexing PCR.
Chromatin coverage Nucleosomal-free regions mapped to > 80% of the genome.
RNA gene coverage > 80% of genes detected as compared to the ted-RNA™ kit for the same sample.
Transcript coverage > 80% of bases from 5’ to 3’ … for mRNA and ncRNA
Ribosomal RNA carryover < 10% without ribosomal removal from total RNA
Reaction volume 10 uL final volume per reaction
Kit components
  1. ted-CapALL™ Enzyme Mix: A proprietary composition of novel enzymes that convert RNA and chromatin into an Illumina compatible sequencing library, in a single tube reaction
  2. ted-CapALL™ Primer Mix: Primers designed to convert RNA and chromatin into an NGS library.
  3. ted-CapALL™ Preamp Oligos: Oligos required to amplify library from ultra low amount to a comfortable level before next step (customer provides PCR reagents).
User provided components
  • Cell lysate or isolated organelles such as exosome
  • SPRI beads system, e.g. AMPure XP or Kapa Pure Bead, etc
  • PCR reagents and indexing adapters (our PCR kit is sold separately)
Applications Simultaneous analysis of
  • chromatin protein occupancy by detection of open regions in the chromosome
  • mRNA, non-polyA coding RNA, non-coding RNA (nc and lncRNA) expression
  • Protocol is compatible with samples from prokaroytic and eukayotic organisms.

Downloads

Simple Workflow

Concordance between total RNA and mRNA input

Gene counts of different amount of input

Reproducibility between different input amounts  (5 to 500 picograms)

Gene coverage 5′ to 3′

Mapped reads categories

Library Structure

X Gen US CO’s products are for Research Use Only. They cannot be used for diagnostic or therapeutic applications. These products cannot be resold, modified for resale, used in the manufacturing of commercial products or transferred to third parties without prior written approval from X Gen US CO. Prior written approval is required for service providers who incorporate X Gen US CO’s products into their service offerings. Prior written approval is required for integrating X Gen US CO’s products into instrumentation systems (automation, liquid handling, cell sorting and related).

X Gen US CO, the logo, ted-RNA, ted-DNA, CapALL and ted-technology are trademarks of X Gen US CO. Illumina is a registered trademark of Illumina, Inc. All other marks are the property of their respective owners. Certain trademarks may not be registered in all jurisdictions.

Additional information

Reactions per kit

8, 96