How do I shear rRNA-depleted RNA?
How do I shear rRNA-depleted RNA?
For cDNA synthesis using the SMARTer Universal Low Input RNA Kit for Sequencing (Cat. # 634938), we recommend shearing at the same time as priming the RNA. To do this, add 4 µl of 5X First-Strand Buffer to your RNA and the 3' SMART N6 CDS Primer II A (not to the Master Mix), then incubate at 94°C. For RNA with a RIN >7, incubate for 5 min. For RNA with RIN 4–7, incubate for 4 min. For RNA with a RIN of 3, incubate for 3 min. For RNA with a RIN <3, follow the protocol as directed in the SMARTer Universal Low Input RNA Kit for Sequencing User Manual.