ted-RNA(™) 5′ WTA Library Preparation Kit

$400.00$3,840.00

mRNA and ncRNA data without rRNA removal
Single tube reaction to convert total RNA or mRNA to Illumina® compatible RNA-seq libraries. < 10% rRNA without rRNA subtraction. 1 to 200 ng total RNA input. For gene expression, RNA isoforms, allele specific expression and other applications for nc-, lnc- and mRNA.
User needs to provide PCR reagents and Illumina® indexing reagents.

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Description

For Illumina sequencing systems

Cat. No. Format Storage Kit stability
TR-1021-08 8 reactions per kit -20°C 1 year from the date of manufacture
TR-1021-96 96 reactions per kit -20°C 1 year from the date of manufacture

The ted-RNA 5′ WTA Library Preparation Kit generates sequencing libraries for Illumina® platforms. The novel “ted” technology used in these kits generate full length transcript coverage from purified total RNA. The method does not require RNA fragmentation before library construction and can generate sequencing libraries with larger insert sizes to produce more accurate RNA isoform data. This kit does not require ribosomal RNA removal as rRNA is converted into library molecules at substantially low efficiency.

Attributes:


*filter duplicated reads to remove rRNA is recommended.
Protocol [PDF] ted-RNA™ 5' WTA Library Preparation Kit User Guide
Input sample 1 - 200 ng total RNA
0.1 - 100 ng mRNA
Data produced Sequencing reads for coding and non-coding RNA with minimal proportion of total reads (< 10%) mapping to ribosomal RNA.
Data processing first 11 bases in Read1 is UMI, Read2 is full length cDNA. Q20 filter and remove duplicates
seqtk trimfq -b 11 $Read1 ; with seqtk from https://github.com/lh3/seqtk.
Protocol time
  1. cDNA library preparation ~30min (1 tube reaction)
  2. Indexing PCR & QC ~1 hr
RNA fragmentation method Not required
Ribosomal removal method Not required
Ribosomal RNA carryover < 10% without ribosomal removal from total RNA*
< 1% from purified mRNA
Transcript coverage > Near full length from Read 2 only; 5' biased from Read 1 only
Reaction volume 10 uL final volume per reaction
Kit components
  1. ted-RNA 5' Enzyme Mix: A proprietary composition of reverse transcriptase and a novel enzyme that converts RNA species into an Illumina compatible sequencing library in a single tube reaction.
  2. ted-RNA 5' Primer Mix: Primers designed to convert RNA molecules to an NGS library with uniform 5’ to 3’ coverage.
  3. Preamp Oligos
User provided components
  • DNA-free total RNA or purified mRNA
  • SPRI beads system, e.g. AMPure XP or Kapa Pure Bead, etc
  • PCR reagents and indexing adapters (our PCR kit is sold separately)
Applications
  • Analysis of mRNA, non-polyA coding RNA, non-coding RNA (nc and lncRNA)
  • Analysis of RNA from prokaryotic or eukaryotic organisms
  • Analysis of RNA isoforms and strand specific transcription

Performance data:

Reproducility & sensitivity
Mapping statistics
IGV Plot
Transcripts Coverage
Genes detected
Library structure Read 1 begins with 11 bases UMI followed by 5' sequence of transcripts

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

XGen, the XGen logo, ted-RNA, ted-DNA, CaptureAll and ted-technology are trademarks of Xgen Laboratories, Inc. 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