The SMARTer Ultra Low RNA Kit for Illumina® Sequencing provides a simple and efficient solution for generating libraries from total RNA that are compatible with Illumina's Genome Analyzer, HiSeq™, and HiScanSQ™ instruments. The kit facilitates transcriptome analysis from as little as 100 pg of input RNA. This highly efficient system for high-throughput RNA sequencing studies allows you to begin with the smallest sample size ever, and end with unparalleled sequencing output.
Integration with Illumina Sequencing
The integration of Clontech’s SMART technology with Illumina sequencing has resulted in the most sensitive sample preparation workflow offered by any next-generation sequencing (NGS) platform (Images & Data tab). The combination of SMART technology’s ability to handle very small quantities of RNA and the Illumina sequencing platform's capacity for single- and paired-end sequencing of millions to billions of long and short reads per run, allows you to annotate coding SNPs, discover transcript isoforms, characterize splice junctions, and determine the relative abundance of transcripts from even the smallest samples.
Unparalleled Sensitivity and High Efficiency
The SMARTer Ultra Low RNA Kit protocol has been specifically developed to improve sensitivity. Now you are able to use a very limited quantity of starting material, such as microdissected tissues, laser-captured cells, biopsy samples, etc. As little as 0.1 ng (100 pg) of total RNA is sufficient for generating a highly representative cDNA pool for library construction and sequencing on Illumina's Genome Analyzer, HiSeq, and HiScanSQ instruments. Although the amount of input RNA can vary over quite a large range (from 1 ng to 0.01 ng), comparable DNA output can be obtained by adjusting the number of PCR cycles. Typical yields of ds cDNA range between 2 ng and 7 ng (Images & Data tab).
Single-Tube Procedure
One of the greatest advantages of SMART (Switching Mechanism at 5’ End of RNA Template) technology is its increased efficiency compared to traditional technologies which require isolation of mRNA and adaptor ligation. The entire SMART protocol is performed by one enzymatic reaction, in a single tube. Your precious RNA is subjected to the least possible handling, minimizing the potential degradation risks.
RNA-seq Data Quality
SMART provides faithfully reproduced, full-length cDNA for use as template in library sample preparation. Sequencing of libraries from mouse brain total RNA at input levels varying from 10 to 0.01 ng demonstrates that even with just 10 pg of input RNA—which is less than the amount found in most single cells—over 90% of the data mapped to the genome, and the average transcript coverage was as uniform as that seen with much greater amounts of RNA (Images & Data tab). Also, under all conditions used, rRNA reads accounted for only 3–5% of the total reads, which is typical for standard poly(A)-selected library preparation methods. All of these results—high mappability, uniform read coverage, number of genes detected, and low amount of rRNA—are entirely consistent with those typically achieved using much larger amounts of RNA. Also, the resulting exon coverage is equivalent to traditional RNA-seq methods requiring significantly more starting material (Images & Data tab).
