How do I lyse cells for direct cDNA synthesis?

Refer to the user manual for the specific kit that you are using, as the lysis reaction conditions may be different for different kits. For the SMART-Seq v4 Ultra Low Input RNA Kit for Sequencing or the SMARTer Ultra Low Input RNA Kit for Sequencing - v3, lysis is conducted at room temperature, while for the previous generations it is done on ice. Lyse the collected cell(s) with Reaction Buffer [...]

What media have been tested for compatibility with direct cDNA synthesis from intact mammalian cells?

It is important to collect cells using media and buffers that do not suppress cDNA synthesis. PBS buffer has been tested and is compatible with all SMARTer Ultra low kits at all inputs (1–9 µl). The following media have not been tested in-house; however, they have been externally validated for use with low-input volumes (1 µl). SuperBlock (Pierce, Cat. # 37515) 0.1 ml of DMEM/F-12, GlutaMAX [...]

Can I use more than 1,000 cells as input for direct cDNA synthesis using SMARTer Ultra low kits?

Using more than 1,000 cells for direct cDNA synthesis with SMARTer Ultra low kits is not recommended. General [...]

How many cells can I use for direct cDNA synthesis with SMARTer Ultra low kits?

SMARTer Ultra low kits can accommodate inputs of 1–1,000 intact mammalian cells for direct cDNA synthesis.Note: Direct cDNA synthesis from plant or insect cells has not been tested in-house with SMARTer Ultra low kits. Electropherogram of cDNA generated from whole cells with the SMARTer Ultra Low Input RNA Kit for Sequencing - v3. General [...]

Which SMARTer Ultra low kit is recommended for direct cDNA synthesis from whole mammalian cells?

For cDNA synthesis from intact cells, we recommend the SMART-Seq v4 Ultra Low Input RNA Kit for Sequencing, as this kit has the highest sensitivity in identifying the maximum number of transcripts from ultra-low inputs of RNA. General [...]