AffiNGS® Discover-sc WTA Kit V2
AffiNGS® Discover-sc WTA Kit V2 is capable of obtaining sufficient samples for sequence analysis through first-strand cDNA synthesis and amplification using 1 - 1,000 cells or 10 pg - 10 ng of total RNA as templates, thus overcoming the technical difficulty that the conventional mRNA-seq method cannot be used for sequence analysis of trace samples such as single cells due to the low RNA content.
The kit employs Oligo dT Primer as the reverse transcription primer for cDNA synthesis. The kit also utilizes the template-switching activity of Discover-sc Reverse Transcriptase to add an adapter sequence at the 3' end of the cDNA, based on which subsequent PCR amplification is performed to obtain full-length cDNA amplification products, effectively avoiding 3' end preferences and rRNA contamination during cDNA synthesis.
Discover-sc WTA Kit V2 is an upgraded version developed on the basis of Discover-sc WTA Kit.
The upgraded version features substantially improved detection sensitivity and volume compatibility, and is more suitable for the detection of low-abundance genes and low-concentration templates. Generally, one reaction can output 2 - 20 ng of cDNA amplification products.
Components
| Components |
AFG-VZ-N711-01 (12 rxns) |
AFG-VZ-N711-02 (24 rxns) |
AFG-VZ-N711-03 (96 rxns) |
|
| BOX 1 | Discover-sc TS Oligo V2 | 12 µl | 24 µl | 96 µl |
| Control Total RNA (1 μg/μl) | 5 µl | 5 µl | 5 µl | |
| BOX 2 | 10X Lysis Buffer V2 | 230 µl | 460 µl | 2 x 920 µl |
| RNase Inhibitor | 20 µl | 40 µl | 160 µl | |
| Oligo dT Primer | 24 µl | 48 µl | 192 µl | |
| dNTP Mix | 24 µl | 48 µl | 192 µl | |
| DTT | 12 µl | 24 µl | 96 µl | |
| 5X FS Buffer V2 | 48 µl | 96 µl | 384 µl | |
| Discover-sc Reverse Transcriptase | 12 µl | 24 µl | 96 µl | |
| 2X Discover-sc PCR Mix | 300 µl | 600 µl | 4 x 600 µl | |
| Discover-sc WTA PCR Primer | 12 µl | 24 µl | 96 µl | |
| Elution Buffer | 1 ml | 1 µl | 2 x 1 ml | |
| Nuclease-free ddH2O | 1 ml | 1 µl | 2 x 1 ml | |
Colored bullets indicate the cap of the reagent to be added.
Storage
BOX 1: Store at -85 ~ -65°C and ship on dry ice.
BOX 2: Store at -30 ~ -15°C and transport at ≤0°C.