TIPS!
Low binding tips, tubes, and plates should always be utilized to ensure the sample doesn’t adhere to the plasticware. This is critical when working with ultra-low inputs and single cells.
For best practice, cells should be kept at 4 °C prior to lysis. Dead or dying cells will result in a decrease in the data quality and may lead to an increase of reads obtained for mitochondrial transcripts.
Step 5: ERE solution to be diluted 1:10 shortly before using (minimum of 8 reactions per experiment).
Step 7: Process to purification immediately after THOR reaction.
Step 13: Proceed to purification immediately after IVT or incubate IVT overnight at 4 °C (or stop and store at -20 °C).