Skip to main content
Skip table of contents

Why do I need to perform a qPCR? Can this step be skipped?

Performing a qPCR assay is recommended to determine the Optimal Endpoint PCR cycle number (OEP). This prevents under- or over-cycling, which may have negative effects on downstream applications. Undercycling results in low yields and often insufficient material for downstream use, while overcycling will result in a loss of longer full-length cDNAs (see Figure). Once the number of cycles for the endpoint PCR is established for a particular RNA sample type (including RNA quality level) and input amount, the same can be used for the following experiments without performing the qPCR each time. See also Appendix B of the TeloPrime V2 User Guide.

Figure | TeloPrime cDNA Quality Control. TeloPrime cDNA was prepared from 500 ng of Universal Human Reference RNA (UHRR). Amplification was performed using 1 ┬Ál of cDNA with 24 cycles of PCR. Final cDNA was run at 1:10 dilution on a Bioanalyzer High Sensitivity DNA Chip (Agilent Technologies). The optimal endpoint PCR (OEP) cycle number was determined by qPCR as described in Appendix B, p.22.

The TeloPrime Full-Length cDNA Amplification Kits (Cat. No. 013) contain sufficient reagents to perform a qPCR and endpoint PCR for each sample. Lexogen also offers a TeloPrime PCR Add-on Kit (Cat. No. 018) for 16 reactions if additional PCRs are required.

JavaScript errors detected

Please note, these errors can depend on your browser setup.

If this problem persists, please contact our support.