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.
Features:
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.
Library quality analysis - basic data index
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.
General analysis of gene expression
|
Sample |
Expression Genes |
Max |
Mean |
1st Qu. |
Median |
3st Qu. |
|||
|
Vazyme-1 |
14807 |
0.002589318 |
5293.111 |
35.207 |
1.020643 |
4.717383 |
20.47274 |
19523.06 |
521310.1 |
|
Vazyme-2 |
15138 |
0.004287681 |
4613 |
35.91853 |
0.954896 |
4.294688 |
18.79261 |
22444.61 |
543734.7 |
|
C-1 |
14367 |
0.004498034 |
4568.409 |
39.20601 |
1.070605 |
4.631338 |
20.44749 |
24545.59 |
563272.8 |
|
C-2 |
15285 |
0.003861415 |
4666.078 |
38.2787 |
0.887525 |
4.007046 |
17.91134 |
26059.52 |
585090 |
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.
Uniform data distribution without preference
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.
High correlation and repeatability of expression
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.