The Single Cell Full Length mRNA Amplification Kit can use 1-500 cells or 10 pg-10 ng of total RNA as a template, using Oligo(dT) VN Primer as a reverse transcription primer for cDNA synthesis, and using the reverse transcription enzyme Sc Reverse Transcriptase.
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Template-switching activity of theSc Reverse Transcriptase enzyme adds a linker sequence to the 3′ end of the cDNA and subsequent PCR amplification of the linker sequence to obtain the full-length cDNA amplification product. The 3′-biase and rRNA over-representation in the cDNA synthesis process is effectively avoided. The sequencing analysis of the obtained full-length cDNA amplification product can enable the retrieval of genetic regulatory information such as gene expression differences, alternative splicing, and fusion genes. Single Cell Full Length mRNA Amplification Kit for compatible sample volume as little as 2.5 μl. In general, one reaction can produce 2-20 ng of cDNA amplification product.
• Low input template: single cells or 10 pg of total RNA can be used as starting template for efficient amplification
• High amplification sensitivity: using the LNA technology and a carefully optimized reaction system , the detection of low-expressed level genes can be accomplished in the library
• Good product integrity: Amplify full-length cDNAs with both forward and reverse primers to obtain full transcriptome information, avoiding 5' and 3' biases.
A single 293T cell and 10 pg Total RNA were amplified for 18 cycles, and 1 μl of the purified cDNA amplification product was tested using an Agilent 2100 Bioanalyzer. The amplified product showed a distribution between 400-10000 bp and the peak size of the library was located at about 2000 bp. No product was observed in the no-template negative control.