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Discover-sc WTA Kit V2
Single Cell Full Length mRNA-Amplification Kit (Catalog # N712)is the recommended replacement for Discover-sc WTA Kit V2(Catalog # N711).
Discover-sc WTA Kit V2
Single Cell Full Length mRNA-Amplification Kit (Catalog # N712) is the recommended replacement for Discover-sc WTA Kit V2(Catalog # N711).
Low starting amount of template: Single cell or 10 pg total RNA can serve as starting templates for efficient amplification.
High sensitivity of amplification: Well optimized reaction system and LNA technology greatly increase the detection rate of low expression genes.
Good integrity of product: Full length cDNA is amplified by two end primers, to obtain information of full transcript, avoiding 5'and 3' preferences.
High success rate of operation: Treatment process of RNA sample is greatly reduced, to eliminate the risk of sample loss to the largest extent, and improve the success rate of operation.
Wide compatibility of volume: The compatible sample volume can be as large as 5 μl, compatible with different concentrations of template for amplification.
Low starting amount of template, and good integrity of cDNA amplification
10 pg total RNA of 293T cells was amplified with Discover-scTM WTA Kit V2, and 1 μl cDNA amplification product was analyzed by Agilent 2100 Bioanalyzer. Oligo dT Primer was used as reverse transcription primers, and a linker sequence was added at the 3 'end of cDNA by Template-switching activity of the Discover-sc Reverse Transcriptase. Following PCR amplification employed the linker sequence, to obtain full-length cDNA amplification products, while avoiding the contamination of rRNA.
Vazyme-1 and Vazyme-2 refer to amplifications of single 293T cell by Discover-scTM WTA Kit V2; C-1 and C-2 refer to amplifications of single 293T cell by single cell transcriptome kit of C company. 1 ng cDNA amplification product were used to construct libraries using TruePrepTM DNA Library Prep Kit V2 for Illumina®. Following information are analyzations of these libraries.
The detection sensitivity of amplification products of Discover-scTM WTA Kit V2 was high, and the number of genes detected in single 293T cell was as high as 15000.
Left picture shows gene distribution results of the Vazyme-1 sequencing fragments, and right picture shows gene distribution results of the C-1 sequencing fragments. Data shows that amplification by Discover-scTM WTA V2 ensures good coverage of gene 5 'and 3' information.
Left picture shows correlation and repeatability analyzation of Vazyme-1 and Vazyme-2, and right picture shows correlation and repeatability analyzation of C-1 and C-2. The result data shows that the correlation and repeatability of our product is higher than that of C company.
Discover-scTM WTA Kit V2 can use total RNA from 1-1000 cells or 10 pg- 10 ng total RNA as template, via first-strand cDNA synthesis and amplification, to achieve enough amount of sample for sequencing analysis, thereby solves the technical problems that sequencing analysis cannot be accomplished by regular mRNA-seq because of low RNA content in small samples such as single cell.
Discover-scTM WTA Kit V2 is an updated version based on Discover-scTM WTA Kit. The detection sensitivity and volume compatibility have been significantly improved, and it is more suitable for detection of templates of low abundance and low concentration.