The SMARTer Ultra Low RNA Kit for Illumina® Sequencing provides a simple and efficient solution for generating libraries for transcriptome analysis (RNA-seq) from single cells or very low amounts of total RNA (10 pg–10 ng) that are compatible with Illumina's Genome Analyzer™, HiSeq®, HiScanSQ™ or MiSeq® instruments. This kit allows you to synthesize cDNA directly from cells or using total RNA as the starting material, simplifying the process of sample preparation. The SMARTer technology (also referred to as SMART-seq) guarantees full length gene coverage that outperforms existing methods in transcriptome analysis, making it an immensely powerful tool for studying single-cell, circulating tumor cells, or alternative splicing events in cancer cells.
Integration with Illumina Sequencing
The SMARTer Ultra Low RNA Kit protocol is compatible with Illumina's Paired-End DNA Sample Prep for preparing libraries to be sequenced on Illumina's sequencing platforms. The SMART technology’s ability to handle very small quantities of RNA, paired with the Illumina sequencing platform's capacity for single- and paired-end sequencing of up 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 sample size.
Unparalleled Sensitivity and High Efficiency
The SMARTer Ultra Low RNA Kit protocol has been specifically developed to provide unparalleled sensitivity to work directly with a single cell, or a few cells. As little as 0.01 ng (10 pg) of total RNA is sufficient for generating a highly representative cDNA pool for library construction and sequencing on Illumina's Genome Analyzer, HiSeq, HiScanSQ or MiSeq instruments. Typical yields of ds cDNA range between 2 ng and 7 ng.
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 risk of degradation.
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, demonstrate that even with just 10 pg of input RNA—which is less than the average amount found in 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. Also, under all conditions tested, 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. Additionally, the resulting exon coverage is equivalent to traditional RNA-seq methods requiring significantly more starting material.
