Small RNAs represent an exciting new frontier in genomics research. The ability to interrogate and quantify small RNAs in a high throughput manner will provide valuable insight into the mechanisms involved in gene regulation and shed new light on the involvement of non-coding RNA in disease states such as cancer.

Helicos allows the interrogation and quantitation of small RNAs without ligation or amplification. Using true single molecule sequencing for small RNA analysis enables the digital profiling of previously identified small RNAsand discovery of novel ones.

The Advantages
Helicos small RNA Sequencing, an amplification-free assay with millions of reads per sample, provides you with:

• The quantitative accuracy only possible with an amplification- and ligation-free approach
• The ability to perform small RNA profiling and discovery on hundreds of samples in a cost-effective manner
• The sequencing accuracy to discover new small RNA species anywhere in the genome
• The resolution to find changes not detectable on array-based platforms or amplification-based next-generation sequencers
• The precision to ensure that your results stay the same from run-to-run and from prep-to-prep

Recent publications utilizing our cDNA-Based Small RNA Sequencing application include:

New class of gene-termini-associated human RNAs suggests a novel RNA copying mechanism. Nature 2010; 466, 642-646.

The Evidence: Virtually unbiased small RNA Quantitation and Discovery
Profiling and discovering small RNAs without amplification or ligation is now a reality. Scientists at Helicos have demonstrated the ability to perform small RNA sequencing experiments using the tSMS approach.

Figure 1. The small RNA fraction (<200nt) was isolated from total RNA using the miRvana™ miRNA Isolation Kit. The small RNA fraction was modified with a poly-C tail using polyA-polymerase. The modified small RNA was converted to cDNA, which was processed using the Helicos DNA Sample Preparation methodology described in figure 1. The modified cDNA sample was loaded into the Helicos Flow Cell, and analyzed using the HeliScope Single Molecule Sequencer. Single molecule reads were mapped to the human genome. Representative results are shown; Black bars: tSMS reads; Blue bars: Known exons; Red bar: Known human miRNA.


The Approach
First-strand cDNA generated from a small RNA fraction is modified with a poly-A tail and loaded onto the instrument. No ligation or PCR amplification steps are required. The tailed fragments hybridize to complementary poly-T strands anchored to the flow cell surface. Inside the HeliScope™ Single Molecule Sequencer, a series of nucleotide addition and detection cycles determine the sequence of each fragment. Open source data analysis software aligns the hundreds of millions of reads to a reference sequence.

Helicos Small RNA Sequencing – See the true small RNA profile of your samples.