How essential is the Thermal Cycler compatibility with the 100ul PCR reaction mix?
How essential is the Thermal Cycler compatibility with the 100ul PCR reaction mix?
It is extremely essential that the Thermal Cycler is compatible with the 100ul PCR reaction mix to ensure precise temperature distribution across the entire sample volume as also indicated in the SMARTer smRNA-Seq Kit for Illumina User Manual - IV. Protocols - C. Protocol: PCR and Cleanup - DO NOT reduce the reaction volume. The 100-µl final volume is important for yield. If your thermal cycler cannot accommodate 100-µl sample volumes, it is important to equally divide each sample into two tubes (containing ~50 µl each) after the PCR Master Mix has been added, mixed, and spun down