Smart-Seq2 Single Cell Kit
mRNA synthesis
In vitro transcription of capped mRNA with modified nucleotides and Poly(A) tail
Tyramide Signal Amplification (TSA)
TSA (Tyramide Signal Amplification), used for signal amplification of ISH, IHC and IC etc.
Phos Binding Reagent Acrylamide
Separation of phosphorylated and non-phosphorylated proteins without phospho-specific antibody
Cell Counting Kit-8 (CCK-8)
A convenient and sensitive way for cell proliferation assay and cytotoxicity assay
SYBR Safe DNA Gel Stain
Safe and sensitive stain for visualization of DNA or RNA in agarose or acrylamide gels.
Inhibitor Cocktails
Protect the integrity of proteins from multiple proteases and phosphatases for different applications.
Smart-Seq2 Single Cell Kit is a highly sensitive cDNA preparation kit that obtains high-quality full-length cDNA from 1-1,000 cells or 10 pg-10 ng total RNA for single cells or trace samples RNA-seq analysis. Smart-Seq2 Single Cell Kit uses Oligo dT30VN primer as the reverse transcription primer to synthesize cDNA, and relies on the template-switching activity of Reverse Transcriptase to add adapter sequence to the 3'-end reverse transcribed full length cDNA. Through subsequent PCR amplification, to obtain full-length cDNA, effectively avoiding the 3' bias and rRNA contamination. The obtained full-length cDNA product can be used for transcriptome sequencing and gene expression analysis.
Smart-Seq2 Single Cell Kit is suitable for the detection of trace samples such as single cells, so that samples with low abundance and low concentration can also be analyzed by RNA-seq sequencing. In general, one reaction can produce 2-20 ng of cDNA amplification products.
Figure 1. Schematic of the technology in the Smart-Seq2 Single Cell Kit
| Components | 12 rxns | 24 rxns | 96 rxns | |
| Box 1 | Control Total RNA* (1 μg/μL) | 5 μL | 5 μL | 5 μL |
| TS Oligo Primer (100 μM) | 2.4 μL | 4.8 μL | 19.2 μL | |
| Box 2 | 10X Lysis Buffer | 230 μL | 460 μL | 920 μL x 2 |
| Oligo dT30VN Primer (10 μM) | 24 μL | 48 μL | 192 μL | |
| dNTP Mixture (10 mM) | 24 μL | 48 μL | 192 μL | |
| 5X First-Strand Buffer | 48 μL | 96 μL | 384 μL | |
| Reverse Transcriptase (200 U/μL) | 12 μL | 24 μL | 96 μL | |
| RNase Inhibitor, Murine (40 U/μL) | 30 μL | 60 μL | 240 μL | |
| Betaine (5 M) | 48 μL | 96 μL | 384 μL | |
| 2X HyPerFUsion® High-Fidelity Master Mix | 300 μL | 600 μL | 1 mL x 3 | |
| IS PCR Primer (10 μM) | 6 μL | 12 μL | 48 μL | |
| Elution Buffer | 1 mL | 1 mL | 1 mL x 2 | |
| Nuclease-free Water | 1 mL | 1 mL | 1 mL x 2 | |
Box 1: Store at -80°C. Box 2: Store at -20°C. *Note: The Control Total RNA is from Hela cell. | ||||
Low starting amount of template：Prepare high-quality full-length cDNA with 1-1,000 cells or 10 pg-10 ng of total RNA template.
High sensitivity of amplification: Combined with Smart-Seq2 and LNA technology, more genes can be detected.
Easy to operate: Single-tube protocol from cell lysis to cDNA, reducing operation steps and operation time.
High success rate of operation: Fewer RNA sample processing steps to avoid the risk of sample loss and increase the success rate of the operation.
