2D Complete RNA
2D Complete RNA
What would you do if you could simultaneously SEE EVERY RNA in your Sample?
Over 1300 unique miRNA and over 19,000 unique protein coding transcripts, along with all other RNA biotypes (including tRNA!) identified using single RNA-Seq library prep.
2D Complete RNA Libraries Provide Unparalleled Library Complexity, and rich data set even at ultra low inputs.
2D Complete RNA Libraries Provides Uniform Coverage Across the Gene Body of Transcripts
Templated used: 10ng Human Universal Reference RNA. Libraries were sequenced on an Illumina HiSeq2500 using single read mode (1x75 bp). This view of the 5´ to 3´coverage of RefSeq transcripts demonstrates consistent and uniform coverage for 2D Complete RNA libraries with 10ng.
Far less PCR cycles required due to high conversion efficiency
About 3-5 cycles less than commonly used methods for similar inputs.
Even Coverage Across High GC Transcripts
Excellent Coverage of Low Abundance Transcripts
Reproducible data across A wide range of input - even in Picogram range
Data from libraries prepared using 2D Complete RNA kit with various input amounts of Human Brain mRNA. Data shows very consistent performance across 3 orders of magnitude of input range. Insets indicates Pearson coefficient of correlation.
FFPE: Robust performance on degraded & compromised sample types
High reproducibility of 2D Complete RNA Kit is confirmed by ERCC analysis.
Reads mapping to ERCC data set from other experiments were pooled together and the FPKM was plotted against relative transcript abundance. All 92 ERCC transcripts were identified.
2D Complete RNA: Molecular Biology & Mechanism of 'Continuous Synthesis'
The Mechanism of “Continuous Synthesis”: 2Dzyme adds the adaptor to cDNA using the mechanism of 'continues synthesis'.
During this process, an oligo dT primer (with partial adaptor) is annealed to the PolyA tail. 2Dzyme performs the reverse transcription reaction creating the cDNA strand till the 5' end of the RNA molecule using its strand-displacement and high-processivity properties. After reaching the 5’ end of the RNA template, 2Dzyme binds second adapter and continues synthesis without relying on the addition of non-templated bases or hybridization to the second adaptor.
The second adaptor is incorporated at high efficiency due to tight/preferential adapter binding by 2Dzyme. The result of this reaction is the cDNA product that includes template and both adapters sequence. More than 99% of the library is automatically stranded due to 2Dzyme's directional reaction. Adapters are engineered to prevent next (third) template binding.
Library Molecule Structure and Sequencing Recommendation:
- Compatible with Illumina Platform
- Dual barcode
- We recommend Sequencing with Single-end reads. This can save on sequencing cost.
- The first 8 bases on Read 2 consist of a random tag sequence that can be used as an Unique Molecular Identifier (UMI). For complete Paired-end reads sequencing, please refer to the FAQ page.
- For Paired-end reads, there can be a significant drop in read quality for read 2 due to the presence of Poly T sequence.
- With 2D Complete RNA kit, single-read sequencing is sufficient to obtain coverage of full transcript.
- During analysis, 1st base should be trimmed before mapping, if it is not soft-clipped.