Lysate starting material

Instructions
 

• Very small amounts of RNA (1-100 cells) can be sequenced directly from cell lysate using the Smart-Seq2 protocol (according to Simone Picelli and Rickard Sandberg et al. Nature Protocols 2014)
• Sort 1 – 100 cells into lysis buffer of a 384 well plate, supplied by the Genomics Unit in the G-INCPM. Sorting should be done under cold condition.
• Before the sorting process, please inform the Genomics Unit regarding the cell volume. The maximum cell volume is 0.825ul.
• At the end of the cell collecting, the plate should undergo quick spin followed by 5-minute incubation at room temperature, followed by immediate freezing at -80 (or dry ice).
• The PCR condition for each cell type has to be determined prior to the real experiment. For that purpose, please bring an additional 12-24 single cells or 5-6 bulk samples. It is also important to add a negative control (lysis buffer without cells).