Do I have to dilute the NTI buffer during purification of the SMARTer smRNA PCR reaction mix?

Follow the standard NucleoSpin PCR Clean-Up and Gel Extraction User Manual protocol without diluting the NTI buffer: 200 ul NTI Buffer is added to 100ul PCR reaction mix.smRNA-Seq [...]

What is the cumulative size of adapters incorporated in the SMARTer smRNA library?

The combined length of 5’ and 3’ library adapters is 153 bp. smRNA-Seq [...]

Can I use AMPure bead purification for PCR -amplfied SMARTer smRNA-Seq instead of NucleoSpin Gel and PCR Clean-Up kit?

We recommend NucleoSpin Gel and PCR Clean-Up kit for the purification of PCR-amplified SMARTer smRNA library because the 100ul PCR reaction volume is rather large , which makes AMPure bead purification challenging and can result in a suboptimal bead: sample separation." ProdCompatibility What is the cumulative size of adapters incorporated in the SMARTer smRNA library? The combined length of 5’ [...]

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 [...]

Do I need to perform small RNA enrichment for compatibility with the SMARTer smRNA-Seq Kit for Illumina?

SMARTer® smRNA-Seq Kit does not require a small RNA enrichment. Total RNA input allows a broader representation of small RNAs.smRNA-Seq [...]