• Analysis of diverse small RNA species: small RNA (15 - 150 nt) sequencing library 제작
• Flexible sample inputs: total RNA 또는 enriched small RNA 1 ng - 2ug
• Illumina®-ready sequencing libraries: Illumina® adaptor와 index를 별도의 Ligation 과정없이 SMARTer® 기술로 부착
• Quick, single-tube workflow: 3시간 이내에 sequencing library 제작 가능

제품설명

Small non-coding RNAs (smRNAs) 발견과 분석 연구는 유전자 발현의 조절 기작을 연구하는 데 있어 큰 기여를 해왔다. 주로 알려진 small RNAs는 miRNAs, siRNAs, piRNAs, snoRNAs로써 20 - 30 nt의 크기를 가지고 있다. SMARTer® smRNA-Seq Kit for Illumina®를 이용하면 소량의 small RNAs (low-input amounts)부터 고품질의 Illumina®-ready sequencing library를 제작 할 수 있다.
본 제품은 1ng - 2ug의 total RNA 또는 enriched small RNA sample로부터 SMART-Seq® v4 기술을 이용하여 Illumina® sequencing library를 제작한다. SMART technology (Switching Mechanism at 5’ End of RNA Template)와 Locked Nucleic Acids (LNAs) 기술이 결합된 SMART-Seq® v4 기술을 이용함으로써, 소량의 small RNAs로부터 편향성이 아주 적은 (minimal bias) NGS library를 제작 가능하다.


그림 1. Schematic of technology used by the SMARTer® smRNA-Seq Kit for Illumina®. SMART technology is used in a ligation-free workflow to generate sequencing libraries for Illumina platforms


그림 2. Demonstrating the accuracy of the SMARTer approach for small RNA-seq. Sequencing libraries were generated from an equimolar pool of 963 synthetic miRNAs (miRXplore Universal Reference) using the SMARTer® smRNA-Seq Kit for Illumina® (1 ng input; purple), or a small RNA-seq kit from a different vendor (Competitor N) employing an adapter ligation method (100 ng input; blue). Following sequencing, mapping, and counting of reads, miRNA expression levels (Y axis, log scale) were normalized, resulting in an expected expression level equal to 1 for each miRNA, and a 2-fold cutoff was assigned both above and below the expected expression level (indicated by two horizontal lines). For visualization purposes, miRNAs are ranked along the X axis in order of expression level.


그림 3. Reproducibility of SMARTer® small RNA-seq data. Sequencing libraries were generated in parallel from the indicated input amounts of human brain total RNA using the SMARTer smRNA-Seq Kit for Illumina, and size selected using the BluePippin system. Following sequencing, data processing, and mapping, expression levels of miRNAs identified for each library were quantified and plotted on correlation diagrams. Panel A. Correlation of miRNA expression levels for experimental replicates involving 1 ng inputs. Panel B. Correlation of miRNA expression levels for 2 μg vs. 1 ng inputs.

Application

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