Can I freeze collected cells prior to cDNA synthesis?
Can I freeze collected cells prior to cDNA synthesis?
If you cannot immediately proceed with cDNA synthesis, you may freeze cells on dry ice and store at –80°C.
Gently centrifuge cells, remove the collection medium and freeze the cell pellets. Collected cells may also be frozen in media compatible with the SMARTer Ultra low protocol. (See ""What media have been tested for compatibility with direct cDNA synthesis from intact mammalian cells?"")
Thaw cells immediately prior to cDNA synthesis and add Reaction Buffer containing RNase Inhibitor.
Allow cell lysis to proceed for 5 minutes at room temperature if using the SMART-Seq v4 Ultra Low Input RNA Kit for Sequencing or the SMARTer Ultra Low Input RNA Kit for Sequencing - v3. If using a legacy kit, lysis can be performed on ice (4°C) or at room temperature.